Substituted pyrimidines as cyclin-dependent kinase inhibitors

ABSTRACT

The present invention discloses a substituted pyridine compound represented by formula I, and a pharmaceutically acceptable salt, stereoisomer and tautomer thereof. The compounds of the present invention are useful in the treatment of cancers.

The present application is a divisional application of U.S. patent application Ser. No. 16/067,158, 371c date Jun. 29, 2018, which is a National Stage of PCT/CN2016/113887, filed Dec. 30, 2016, which claims priority benefit of Chinese Patent Application No. CN201511028799.6, filed Dec. 31, 2015, Chinese Patent Application No. CN201610516637.5, filed Jul. 1, 2016, and Chinese Patent Application No. CN201610877404.8, filed Sep. 30, 2016. Each application is hereby incorporated herein by reference in its entirety.

FIELD

The present invention relates to a nitrogen-containing fused heterocyclic compound, as well as a preparation method, an intermediate, a composition and a use thereof.

BACKGROUND

Tumor is a type of cell cycle disease (CCD). Regulating or blocking the cell cycle is one of the ways to treat tumor. At present, many molecules involved in the cell cycle regulation have been discovered, wherein Cyclin-Dependent-Kinases (CDKs) are the core molecules in the cell cycle regulatory network. CDKs are catalytic subunits and are a family of serine (Ser)/threonine (Thr) kinases, they function as important signal transduction molecules in cells, form CDK-cyclin complexes with cyclins and are involved in cell growth, proliferation, dormancy, or entry into apoptosis. Cell cycle regulatory proteins play an important role in the regulation of cell proliferation. In the tumor cells, G1 cyclins and CDK disorders are the most frequent variations in which many mechanisms are involved. These variations often cause tumorigenesis by activating tumor genes and silencing tumor suppressor genes. Malignant cells affect the expression of cell cycle regulatory proteins through genetic and growing mechanisms, causing overexpression of cyclins and expression loss of CDK inhibitors followed by incontrollable CDK activity.

Cell cycle checkpoints are the rate-limiting sites that control cell proliferation cycle, responsible for determining the integrity of DNA synthesis, monitoring DNA replication, repairing DNA damage, and blocking entrance into mitosis period before DNA replication and mitosis, they precisely regulate the progression of cell cycle and prevent errors in the proliferation cycle. Cells respond to DNA damage, which activates the cell cycle checkpoints and cause cell cycle arrest to repair damaged DNA, or induce cell death by apoptosis or termination of growth. Cell cycle arrest often occurs at the G1/S or G2/M junction, and the cell cycle checkpoint regulatory molecules are also new targets for antitumor drugs.

In the development of drugs, the first generation of CDK inhibitors represented by flavopiridol, UCN-01 etc. are “pan-CDK” inhibitors, which blocks all the subtypes of the CDK family in an equivalent manner, exhibits relatively high toxicity in clinical trials and some of them cannot reach the effective therapeutic dose. Therefore, people are inspired to develop selective CDK inhibitors, which are expected to improve the selectivity of treatment and prevent normal cells from damages of side effects. In recent years, selective CDK inhibitors have been reported and entered clinical trials. Therefore, discovering highly specific and low-toxic cell cycle regulators is a frontier in the research of antitumor drugs.

Content

The technical problem to be solved in the present invention is to overcome the existing cell cycle regulators accompanied with severe clinical side effects due to low specificity, poor selectivity etc., or poor activity in spite of high selectivity. Thus, the present invention provides a nitrogen-containing fused heterocyclic compound, as well as a preparation method, an intermediate, a composition and a use thereof, the compounds exhibit a high selectivity and a high inhibitory activity with respect to CDK4 and CDK6 at a molecular level, an excellent inhibitory activity with respect to breast cancer cells at a cellular level, and significant inhibition of tumor cell proliferation associated with cyclin-dependent kinase activity at an animal level. The compounds also exhibits a good stability with respect to human or mouse liver microsomes without significant inhibition of metabolic enzymes, good in vivo absorption in mice and rats, a high bioavailability and good druggability.

The present invention provides a nitrogen-containing fused heterocyclic compound represented by formula (I), a pharmaceutically acceptable salt thereof, an enantiomer thereof, a diastereomer thereof, a tautomer thereof, a solvate thereof, an metabolite thereof or a drug precursor thereof;

wherein, p and q are independently 0 or 1;

m and n are independently 0, 1 or 2;

X and Y are independently CH or N;

the N atom in the cycle

is oxidized or non-oxidized;

preferably, at least one of p and q is 1 (for example, when p and q are both 1, m is 0, 1 or 2, n is 0; or, when p and q are both 1, n is 0, 1 or 2, m is 0; or, when p is 0, q is 1, m and n are independently 0, 1 or 2; or, when p is 1, q is 0, m and n are independently 0, 1 or 2);

R¹ is hydrogen, hydroxy, cyano, thio, substituted or unsubstituted C₂-C₂₀ alkenyl, substituted or unsubstituted C₂-C₂₀ alkynyl, halogen (e.g. fluorine), substituted or unsubstituted C₁-C₂₀ alkyl (e.g. methyl, trifluoromethyl), C₁-C₂₀ alkylthio, substituted or unsubstituted C₁-C₂₀ alkoxy (e.g. methoxy), substituted or unsubstituted C₃-C₁₂ cycloalkyl (e.g. cyclopropyl), substituted or unsubstituted C₁-C₉ heterocycloalkyl, or “C₁-C₂₀ alkyl mono-substituted, di-substituted or unsubstituted amino”; wherein, the two substituents in the “C₁-C₂₀ alkyl di-substituted amino” are the same or different;

R² is hydrogen, hydroxy, substituted or unsubstituted C₂-C₂₀ alkenyl, substituted or unsubstituted C₂-C₂₀ alkynyl, halogen, substituted or unsubstituted C₁-C₂₀ alkyl (e.g. methyl), substituted or unsubstituted C₁-C₂₀ alkoxy (e.g. methoxy), substituted or unsubstituted C₁-C₂₀ alkylthio (e.g. methylthio), substituted or unsubstituted C₃-C₁₂ cycloalkyl (e.g. cyclopropyl), cyano, “C₁-C₂₀ alkyl mono-substituted, di-substituted or unsubstituted amino” or

wherein, the two substituents in the “C₁-C₂₀ alkyl di-substituted amino” are the same or different;

R³ is hydrogen, substituted or unsubstituted C₁-C₂₀ alkyl (e.g. methyl, ethyl, isopropyl, tert-butyl, 1-ethylpropyl, 1-cyclopropylethyl, 2-fluoroethyl, 2,2-difluoroethyl,2,2,2-trifluoroethyl,1-methyl-2,2,2-trifluoroethylorcyclopropylmethyl), substituted or unsubstituted C₂-C₂₀ alkenyl, substituted or unsubstituted C₂-C₂₀ alkynyl, substituted or unsubstituted C₃-C₁₂ cycloalkyl (e.g. 1-methylcyclopropyl, cyclopropyl, 3-oxocyclobutyl 3-hydroxycyclobutyl, 3-fluorocyclobutyl, 3,3-difluorocyclobutyl, cyclobutyl, cyclopentyl, cyclohexyl, or

or substituted or unsubstituted C₁-C₉ heterocycloalkyl (e.g. C₃-C₅ heterocycloalkyl, preferably C₃ heterocycloalkyl; e.g. 3-oxetanyl, 3-methyl-3-azacyclobutyl); wherein, the “heterocycloalkyl” connects to another group via the carbon atom thereof, the heteroatom in the heterocycloalkyl is selected from the group consisting of O, N and S, the number of the heteroatom is 1 to 3, when the heteroatom is N, N can be further substituted with C₁-C₃ alkyl, the heteroatom can be located at the para-position of the carbon atom that connects to another group (e.g. R³ is

the “substituted” in the “substituted or unsubstituted C₁-C₂₀ alkyl”, “substituted or unsubstituted C₂-C₂₀ alkenyl”, “substituted or unsubstituted C₂-C₂₀ alkynyl”, “substituted or unsubstituted C₁-C₂₀ alkoxy”, “substituted or unsubstituted C₁-C₉ heterocycloalkyl”, “substituted or unsubstituted C₃-C₁₂ cycloalkyl” and “substituted or unsubstituted C₁-C₂₀ alkylthio” in the definitions of R¹, R² and R³ refers to be independently substituted with the substituent selected from the group consisting of halogen (e.g. fluorine, the number can be 1, 2 or 3), hydroxyl, oxo (e.g. an oxo group attached to C₃, C₄ or C₅ cycloalkyl, e.g.

thio, cyano, amino, nitro, C₁-C₂₀ alkyl (e.g. methyl), C₂-C₂₀ alkenyl, C₂-C₂₀ alkynyl, C₁-C₂₀ alkoxy, C₁-C₂₀ alkylthio, C₃-C₁₂ cycloalkyl (e.g. cyclopropyl) and “C₁-C₂₀ alkyl mono-substituted, di-substituted or unsubstituted amino”; wherein, the two substituents in the “C₁-C₂ alkyl di-substituted amino” are the same or different; when there are more than one substituents, the substituents are the same or different;

R⁴ is hydrogen, halogen (e.g. fluorine), C₁-C₆ alkyl, C₁-C₃ alkoxy (e.g. methoxy), cyano or

R⁷ contained in R² and R⁴ is independently hydrogen, hydroxyl, C₁-C₆ alkyl (e.g. methyl or ethyl), substituted or unsubstituted C₃-C₈ cycloalkyl (e.g. cyclobutyl, cyclopropyl or 4-methylcyclohexyl), C₁-C₆ alkoxy or “C₁-C₆ alkyl mono-substituted, di-substituted or unsubstituted amino”; wherein, the “substituted” in the “substituted or unsubstituted C₃-C₈ cycloalkyl” refers to be substituted by C₁-C₆ alkyl; the two substituents in the “C₁-C₆ alkyl di-substituted amino” are the same or different;

at least one of R¹ and R⁴ is halogen;

R⁵ is hydrogen, substituted or unsubstituted C₁-C₂₀ alkyl (e.g. substituted or unsubstituted C₁-C₆ alkyl, preferably substituted or unsubstituted methyl, ethyl or propyl; e.g. substituted or unsubstituted n-propyl or isopropyl; e.g. substituted or unsubstituted 2-methylpropyl; e.g. substituted or unsubstituted 1,2-dimethylpropyl; e.g. 2-methoxyethyl, 2-hydroxyethyl, 2-fluoroethyl, 2,2,2-trifluoroethyl, dimethylaminomethyl or 2-dimethylamino-ethyl), substituted or unsubstituted C₂-C₂₀ alkenyl (e.g. vinyl), substituted or unsubstituted C₂-C₂₀ alkynyl (e.g. ethynyl, 2-butynyl or 2-propynyl), substituted or unsubstituted C₃-C₁₂ cycloalkyl

substituted or unsubstituted C₁-C₉ heterocycloalkyl (e.g. tetrahydrogenpyrrolyl,

substituted or unsubstituted C₁-C₉ heteroaryl, substituted or unsubstituted C₃-C₁₂ aryl,

wherein the “C₁-C₉ heterocycloalkyl” connects to another group via the carbon atom thereof;

in R⁵, each R⁸ is independently hydroxyl, substituted or unsubstituted C₁-C₂₀ alkyl (the “substituted or unsubstituted C₁-C₂₀ alkyl” is preferably “substituted or unsubstituted C₁-C₆ alkyl”, preferably “substituted or unsubstituted methyl or ethyl”; e.g. dimethylaminomethyl), substituted or unsubstituted C₂-C₂₀ alkenyl (e.g. vinyl), substituted or unsubstituted C₂-C₂₀ alkynyl (e.g. ethynyl), substituted or unsubstituted C₃-C₁₂ cycloalkyl, substituted or unsubstituted C₁-C₉ heteroaryl, substituted or unsubstituted C₃-C₁₂ aryl, substituted or unsubstituted C₁-C₉ heterocycloalkyl (e.g.

substituted or unsubstituted C₁-C₂₀ alkoxy (e.g. methoxy or ethoxy) or

the “C₁-C₉ heterocycloalkyl” connects to another group via a carbon atom or a heteroatom thereof;

in R⁵, each R⁹ is independently hydrogen, hydroxyl, substituted or unsubstituted C₁-C₂₀ alkyl (e.g. methyl, ethyl, isopropyl, aminomethyl, hydroxymethyl, hydroxyethyl, methylaminomethyl, dimethylaminomethyl, 2-dimethylamino-ethyl,

or methoxymethyl; the “C₁-C₂₀ alkyl” can be methyl, ethyl,

substituted or unsubstituted C₂-C₂₀ alkenyl (e.g. vinyl), substituted or unsubstituted C₂-C₂₀ alkynyl (e.g. ethynyl), substituted or unsubstituted C₃-C₁₂ cycloalkyl (e.g. cyclobutyl,

or 4-methylcyclohexyl), substituted or unsubstituted C₁-C₂₀ alkoxy (e.g. 2-dimethylamino-ethoxy; the “C₁-C₂₀ alkoxy” can be methoxy or ethoxy), substituted or unsubstituted C₃-C₁₂ aryl, substituted or unsubstituted C₁-C₉ heteroaryl, or substituted or unsubstituted C₁-C₉ heterocycloalkyl

the “C₁-C₉ heterocycloalkyl” connects to another group via a carbon atom or a heteroatom thereof;

in R⁸ and R⁹, each R^(10a) and R^(10b) is independently hydrogen, halogen, substituted or unsubstituted C₁-C₂₀ alkyl (e.g. 2-dimethylamino-ethyl or 2-hydroxyethyl; wherein the “C₁-C₂₀ alkyl” can be ethyl), substituted or unsubstituted C₂-C₂₀ alkenyl, substituted or unsubstituted C₂-C₂₀ alkynyl, substituted or unsubstituted C₃-C₁₂ cycloalkyl, substituted or unsubstituted C₁-C₉ heteroaryl, substituted or unsubstituted C₃-C₁₂ aryl, substituted or unsubstituted C₁-C₉ heterocycloalkyl

wherein the “C₁-C₉ heterocycloalkyl” connects to another group via a carbon atom or a heteroatom thereof;

the “substituted” in the “substituted or unsubstituted C₁-C₂₀ alkyl”, “substituted or unsubstituted C₂-C₂₀ alkenyl”, “substituted or unsubstituted C₂-C₂₀ alkynyl”, “substituted or unsubstituted C₃-C₁₂ cycloalkyl”, “substituted or unsubstituted C₁-C₉ heterocycloalkyl”, “substituted or unsubstituted C₃-C₁₂ aryl”, “substituted or unsubstituted C₁-C₉ heteroaryl”, “substituted or unsubstituted C₁-C₂₀ alkylthio” and “substituted or unsubstituted C₁-C₂₀ alkoxy” in the definitions of R⁵, R⁸, R⁹, R^(10a) and R^(10b) refers to be independently substituted with the substituent selected from the group consisting of

C₁-C₂₀ alkyl (e.g. methyl, isopropyl), C₂-C₂₀ alkenyl (e.g. vinyl), C₂-C₂₀ alkynyl (e.g. 2-propynyl), hydroxy, nitro, carboxyl, formyl, thio, trifluoromethyl, C₁-C₂₀ silyl

wherein R^(si1), R^(si2), R^(si3) are each independently C₁-C₃ alkyl such as methyl), halogen (e.g. fluorine, the number can be 1, 2 or 3), cyano, C₁-C₂₀ alkoxy (e.g. methoxy), C₃-C₁₂ cycloalkyl, substituted or unsubstituted C₁-C₉ saturated or unsaturated heterocycloalkyl (the “substituted” refers to be independently substituted with the substituent selected from the group consisting of methylene, oxo, thio, halogen, trifluoromethyl, hydroxy, —PO(OH)₂, —PO(OR^(ps))₂, C₁-C₂₀ alkyl which is optionally substituted with propynyl, C₁-C₂₀ alkyl which is optionally substituted with ethynyl, C₁-C₆ alkenyl, C₁-C₂₀ alkynyl, —B(OH)₂ and —B(OR^(bs))₂, wherein R^(ps) and R^(bs) are independently C₁-C₆ alkyl such as methyl, ethyl or propyl, the C₁-C₂₀ alkyl can be C₁-C₆ alkyl such as methyl, ethyl or propyl; unsaturated heterocycloalkyl such as

saturated heterocycloalkyl such as

when there are more than one substituents, the substituents are the same or different),

thioureido C₃-C₁₂ aryl, C₁-C₉ heteroaryl,

—SeR¹⁹ and —SF₅; wherein heteroatom can be oxidized, N atom can be quaternized; wherein the “heterocycloalkyl” connects to another group via a carbon atom or a heteroatom thereof, the N atom in

isoxidized or non-oxidized, when there are more than one substituents, the substituents are the same or different;

wherein, R^(11a) and R^(11b) are independently hydrogen, halogen, substituted or unsubstituted C₁-C₂₀ alkyl (e.g. substituted or unsubstituted C₁-C₆ alkyl, preferably substituted or unsubstituted C₁, C₂ or C₃ alkyl; e.g. —CF₃; e.g. hydroxyethyl, 2-dimethylaminoethyl), substituted or unsubstituted C₂-C₂₀ alkenyl (e.g. 2-propenyl), substituted or unsubstituted C₂-C₂₀ alkynyl (e.g. 2-propynyl), substituted or unsubstituted C₃-C₁₂ cycloalkyl, substituted or unsubstituted C₁-C₉ heterocycloalkyl

the substituent such as “C₁-C₂₀ alkyl”), guanidino, thioureido, substituted or unsubstituted C₃-C₁₂ aryl substituted or unsubstituted C₁-C₉ heteroaryl,

−SeR²⁶ or —SF₅; wherein the heteroatom can be oxidized, N atom can be quaternized; the “heterocycloalkyl” connects to another group via a carbon atom or a heteroatom thereof, or, in

R^(11a) and R^(11b) together with the N atom attached form

wherein t and u are independently 1, 2 or 3, R^(c) is C(R^(cs))₂, NR^(cs), O, S, Si(R^(cs))₂ or Se, each R^(cs) is independently H, C₁-C₆ alkyl (e.g. methyl, ethyl, propyl or isopropyl), or B(OH)₂; or, in

R^(11a) and R^(11b) together with the N atom attached form

wherein v and w are independently 1, 2 or 3, R^(ch) is CH or PH₂, R^(cx) is CH₂, O, S or ═N—OH, R^(cx) is preferably attached to R^(ch)

R^(hc) is H or C₁-C₆ alkyl (e.g. methyl, ethyl or propyl), preferably when R^(ch) is P, R^(hc) is C₁-C₆ alkyl; or in

R^(11a) and R^(11b) together with the N atom attached form

wherein v1 and w1 are independently 1, 2 or 3, R^(c1) and R^(c2) are independently O, NH or N—CN;

wherein the “substituted” in the “substituted or unsubstituted C₁-C₂₀ alkyl”, “substituted or unsubstituted C₂-C₂₀ alkenyl”, “substituted or unsubstituted C₂-C₂₀ alkynyl”, “substituted or unsubstituted C₃-C₁₂ cycloalkyl”, “substituted or unsubstituted C₁-C₉ heterocycloalkyl”, “substituted or unsubstituted C₃-C₁₂ aryl” and “substituted or unsubstituted C₁-C₉ heteroaryl” in the definitions of R^(11a) and R^(11b) refers to be independently substituted with the substituent selected from the group consisting of C₁-C₂₀ alkyl, C₂-C₂₀ alkenyl (e.g. vinyl), C₂-C₂₀ alkynyl (e.g. ethynyl), hydroxy, nitro, carboxyl, formyl, thio, C₁-C₂₀ silyl

wherein R^(si1), R^(si2), R^(si3) are each independently C₁-C₃ alkyl such as methyl), halogen,

trifluoromethyl, cyano, C₁-C₂₀ alkoxy, C₃-C₁₂ cycloalkyl, C₁-C₉ heterocycloalkyl, C₃-C₁₂ aryl, C₁-C₉ heteroaryl,

and —SF₅, when there are more than one substituents, the substituents are the same or different; wherein the heteroatom can be oxidized, N atom can be quaternized; wherein the “heterocycloalkyl” connects to another group via a carbon atom or a heteroatom thereof;

wherein each R¹² is independently hydrogen, halogen, C₁-C₂₀ alkyl (e.g. methyl), C₂-C₂₀ alkenyl, C₂-C₂₀ alkynyl, hydroxy, —CF₃, —SF₅, C₁-C₂₀ alkoxy (e.g. methoxy or ethoxy) or

wherein R¹³ is hydrogen, C₁-C₂₀ alkyl (e.g. methyl), C₂-C₂₀ alkenyl, C₂-C₂₀ alkynyl, hydroxy, —CF₃, —SF₅ or

wherein each R¹⁴ is independently halogen, C₁-C₂₀ alkyl, C₂-C₂₀ alkenyl, C₂-C₂₀ alkynyl, hydroxy, —CF₃, —SF₅, C₁-C₂₀ alkoxy or

wherein each R¹⁵, R^(16a) and R^(16b) is independently hydrogen, halogen, hydroxy, cyano, nitro, amino, formyl, C₁-C₂₀ alkoxy, C₁-C₂₀ alkylthio, C₁-C₂₀ alkyl (e.g. methyl), C₂-C₂₀ alkenyl, C₂-C₂₀ alkynyl,

—CF₃, —SF₅, C₃-C₁₂ cycloalkyl, or “C₁-C₉ heterocycloalkyl which is optionally substituted with C₁-C₂₀ alkyl”

wherein the “C₁-C₉ heterocycloalkyl” connects to another group via a carbon atom or a heteroatom thereof;

wherein R^(17a) is independently hydrogen, halogen, hydroxy, cyano, nitro, amino, formyl, C₁-C₂₀ alkoxy, C₁-C₂₀ alkylthio, C₁-C₂₀ alkyl (e.g. methyl), C₂-C₂₀ alkenyl, C₂-C₂₀ alkynyl, —CF₃, —SF₅, —SeR²⁶,

C₃-C₁₂ cycloalkyl or “C₁-C₉ heterocycloalkyl which is optionally substituted with C₁-C₂₀ alkyl”

wherein the “C₁-C₉ heterocycloalkyl” connects to another group via a carbon atom or a heteroatom thereof;

wherein R^(17b) is independently hydrogen, halogen, hydroxy, cyano, nitro, amino, formyl, C₁-C₂₀ alkoxy, C₁-C₂₀ alkyl, C₂-C₂₀ alkenyl, C₂-C₂₀ alkynyl, —CF₃, —SF₅,

−OR²⁸, C₃-C₁₂ cycloalkyl or “C₁-C₉ heterocycloalkyl which is optionally substituted with C₁-C₂₀ alkyl”, wherein the “C₁-C₉ heterocycloalkyl” connects to another group via a carbon atom or a heteroatom thereof;

or, R^(17a) and R^(17b) together with the carbon atom, N atom

attached form substituted or unsubstituted C₁-C₉ heterocycloalkyl; wherein the “substituted” in the “substituted or unsubstituted C₁-C₉ heterocycloalkyl” refers to be independently substituted with the substituent selected from the group consisting of C₁-C₂₀ alkyl, C₂-C₂₀ alkenyl (e.g. vinyl), C₂-C₂₀ alkynyl (e.g. ethynyl), hydroxy, nitro, carboxyl, formyl, thio, C₁-C₂₀ silyl

wherein R^(si1), R^(si2), R^(si3) are each independently C₁-C₃ alkyl such as methyl), halogen,

trifluoromethyl, cyano, C₁-C₂₀ alkoxy, C₃-C₁₂ cycloalkyl, C₁-C₉ heterocycloalkyl, C₃-C₁₂ aryl, C₁-C₉ heteroaryl,

and −SF₅, when there are more than one substituents, the substituents are the same or different; wherein the “substituted or unsubstituted C₁-C₉ heterocycloalkyl” connects to another group via a carbon atom or a heteroatom thereof;

wherein R^(18a) and R^(18b) are independently hydrogen, halogen, hydroxy, cyano, C₁-C₂₀ alkoxy, C₁-C₂₀ alkyl, C₂-C₂₀ alkenyl, C₂-C₂₀ alkynyl, —CF₃, —SF₅, C₃-C₁₂ cycloalkyl or “C₁-C₉ heterocycloalkyl which is optionally substituted with C₁-C₂₀ alkyl”; wherein the “C₁-C₉ heterocycloalkyl” connects to another group via a carbon atom or a heteroatom thereof;

or, R^(18a) and R^(18b) together with the heteroatom attached form substituted or unsubstituted C₁-C₉ heterocycloalkyl; wherein the “substituted” in the “substituted or unsubstituted C₁-C₉ heterocycloalkyl” refers to be independently substituted with the substituent selected from the group consisting of C₁-C₂₀ alkyl, C₂-C₂₀ alkenyl (e.g. vinyl), C₂-C₂₀ alkynyl (e.g. ethynyl), hydroxy, nitro, carboxyl, formyl, thio, C₁-C₂₀ silyl

wherein R^(si1), R^(si2), R^(si3) are each independently C₁-C₃ alkyl such as methyl), halogen,

trifluoromethyl, cyano, C₁-C₂₀ alkoxy, C₃-C₁₂ cycloalkyl, C₁-C₉ heterocycloalkyl, C₃-C₁₂ aryl, C₁-C₉ heteroaryl,

and −SF₅, when there are more than one substituents, the substituents are the same or different; wherein the “C₁-C₉ heterocycloalkyl” connects to another group via a carbon atom or a heteroatom thereof;

wherein R¹⁹ is C₁-C₂₀ alkyl, C₂-C₂₀ alkenyl, C₂-C₂₀ alkynyl, C₃-C₁₂ cycloalkyl, C₁-C₉ heterocycloalkyl, C₃-C₁₂ aryl, C₁-C₉ heteroaryl;

wherein the R²⁰ is independently hydrogen, halogen, C₁-C₂₀ alkyl, C₂-C₂₀ alkenyl, C₂-C₂₀ alkynyl, hydroxy, —CF₃, —SF₅, C₃-C₅ heteroaryl (wherein the number of heteroatom is 1 to 3, the heteroatom is O, N or S,

C₁-C₂₀ alkoxy or

wherein R²¹ in the definitions of R^(11a) and R^(11b) is independently halogen, C₁-C₂₀ alkyl, C₂-C₂₀ alkenyl, C₂-C₂₀ alkynyl, hydroxy, —CF₃, —SF₅, C₁-C₂₀ alkoxy or

wherein the R²², R^(23a) and R^(23b) are independently hydrogen, halogen, hydroxy, cyano, nitro, amino, formyl, C₁-C₂₀ alkoxy, C₁-C₂₀ alkylthio, C₁-C₂₀ alkyl, C₂-C₂₀ alkenyl, C₂-C₂₀ alkynyl,

—CF₃, —SF₅, C₃-C₁₂ cycloalkyl, or “C₁-C₉ heterocycloalkyl which is optionally substituted with C₁-C₂₀ alkyl”; wherein the “C₁-C₉ heterocycloalkyl” connects to another group via a carbon atom or a heteroatom thereof;

wherein R^(24a) is independently hydrogen, halogen, hydroxy, cyano, nitro, amino, formyl, C₁-C₂₀ alkoxy, C₁-C₂₀ alkylthio, C₁-C₂₀ alkyl(e.g. methyl), C₂-C₂₀ alkenyl, C₂-C₂₀ alkynyl, —CF₃, —SF₅,

—SeR³¹, C₃-C₁₂ cycloalkyl or “C₁-C₉ heterocycloalkyl which is optionally substituted with C₁-C₂₀ alkyl”; wherein the “C₁-C₉ heterocycloalkyl” connects to another group via a carbon atom or a heteroatom thereof;

wherein R^(24b) is independently hydrogen, halogen, hydroxy, cyano, nitro, amino, formyl, C₁-C₂₀ alkoxy, C₁-C₂₀ alkyl(e.g. methyl), C₂-C₂₀ alkenyl, C₂-C₂₀ alkynyl, —CF₃, —SF₅,

—OR³², C₃-C₁₂ cycloalkyl or “C₁-C₉ heterocycloalkyl which is optionally substituted with C₁-C₂₀ alkyl”

wherein the “C₁-C₉ heterocycloalkyl” connects to another group via a carbon atom or a heteroatom thereof;

or, R^(24a) and R^(24b) together with the carbon atom, N atom attached form substituted or unsubstituted C₁-C₉ heterocycloalkyl; wherein the “substituted” in the “substituted or unsubstituted C₁-C₉ heterocycloalkyl” refers to be independently substituted with the substituent selected from the group consisting of halogen, hydroxy, cyano, nitro, amino, formyl, carboxyl, thio, C₁-C₂₀ silyl, C₁-C₂ alkoxy, C₁-C₂ alkyl(e.g. methyl), C₂-C₂₀ alkenyl(e.g. vinyl), C₂-C₂₀ alkynyl(e.g. ethynyl), —CF₃, —SF₅,

C₃-C₁₂ cycloalkyl, C₁-C₉ heterocycloalkyl, C₃-C₁₂ aryl, C₁-C₉ heteroaryl,

—SeR³¹, “C₁-C₂₀ alkyl mono-substituted, di-substituted or unsubstituted amino” and —SF₅, when there are more than one substituents, the substituents are the same or different; wherein the “substituted or unsubstituted C₁-C₉ heterocycloalkyl” connects to another group via a carbon atom or a heteroatom thereof;

wherein the R^(25a) and R^(25b) are independently hydrogen, halogen, hydroxy, cyano, C₁-C₂₀ alkoxy(e.g. ethoxy), C₁-C₂₀ alkyl(e.g. methyl), C₂-C₂₀ alkenyl, C₂-C₂₀ alkynyl, —CF₃, —SF₅, C₃-C₁₂ cycloalkyl or “C₁-C₉ heterocycloalkyl which is optionally substituted with C₁-C₂₀ alkyl”

wherein the “C₁-C₉ heterocycloalkyl” connects to another group via a carbon atom or a heteroatom thereof;

or, R^(25a) and R^(25b) together with the heteroatom attached form substituted or unsubstituted C₁-C₉ heterocycloalkyl; wherein the “substituted” in the “substituted or unsubstituted C₁-C₉ heterocycloalkyl” refers to be independently substituted with the substituent selected from the group consisting of halogen, hydroxy, cyano, nitro, amino, formyl, carboxyl, thio, C₁-C₂₀ silyl, C₁-C₂₀ alkoxy, C₁-C₂₀ alkyl(e.g. methyl), C₂-C₂₀ alkenyl (e.g. vinyl), C₂-C₂₀ alkynyl(e.g. ethynyl), —CF₃, —SF₅,

C₃-C₁₂ cycloalkyl, C₁-C₉ heterocycloalkyl, C₃-C₁₂ aryl C₁-C₉ heteroaryl,

—SeR³¹, “C₁-C₂₀alkyl mono-substituted, di-substituted or unsubstituted amino” and —SF₅, when there are more than one substituents, the substituents are the same or different; wherein the “C₁-C₉ heterocycloalkyl” connects to another group via a carbon atom or a heteroatom thereof;

wherein R²⁶ is independently C₁-C₂₀ alkyl, C₂-C₂₀ alkenyl, C₂-C₂₀ alkynyl, C₃-C₁₂ cycloalkyl, C₁-C₉ heterocycloalkyl, C₃-C₁₂ aryl or C₁-C₉ heteroaryl;

wherein R^(27a) and R^(27b) are independently hydrogen, halogen, hydroxy, cyano, formyl, C₁-C₂₀ alkoxy, C₂-C₂₀ alkenyl, C₂-C₂₀ alkynyl, —CF₃, —SF, C₃-C₁₂ cycloalkyl, substituted or unsubstituted C₁-C₉ heterocycloalkyl

or substituted or unsubstituted C₁-C₂₀ alkyl(e.g. C₁-C₆ alkyl, preferably methyl, ethyl or propyl); wherein the “substituted” in the “substituted or unsubstituted C₁-C₂₀ alkyl” and “substituted or unsubstituted C₁-C₉ heterocycloalkyl” refers to be independently substituted with the substituent selected from the group consisting of halogen, C₁-C₂₀ alkoxy, C₁-C₂₀ alkylthio and C₁-C₂₀ silyl, when there are more than one substituents, the substituents are the same or different; wherein the “C₁-C₉ heterocycloalkyl” connects to another group via a carbon atom or a heteroatom thereof;

wherein R²⁸ is independently C₁-C₂₀ alkyl, C₂-C₂₀ alkenyl, C₂-C₂₀ alkynyl, or substituted or unsubstituted C₁-C₂₀ silyl; wherein the substituent in the “substituted or unsubstituted C₁-C₂₀ silyl” is C₂-C₂₀ alkenyl or C₂-C₂₀ alkynyl;

wherein R^(29a) and R^(29b) are independently hydrogen, halogen, hydroxy, cyano, formyl, C₁-C₂₀ alkoxy, C₂-C₂₀ alkenyl, C₂-C₂₀ alkynyl, —CF₃, —SF₅, C₃-C₁₂ cycloalkyl, substituted or unsubstituted C₁-C₉ heterocycloalkyl

or substituted or unsubstituted C₁-C₂₀ alkyl; wherein the “substituted” in the “substituted or unsubstituted C₁-C₂₀ alkyl” and “substituted or unsubstituted C₁-C₉ heterocycloalkyl” refers to be independently substituted with the substituent selected from the group consisting of halogen, C₁-C₂₀ alkoxy, C₁-C₂₀ alkylthio and C₁-C₂₀ silyl, when there are more than one substituents, the substituents are the same or different; wherein the “C₁-C₉ heterocycloalkyl” connects to another group via a carbon atom thereof;

wherein R³⁰ is independently hydrogen, hydroxy, C₁-C₂₀ alkyl which is optionally substituted with halogen, or C₁-C₂₀ alkyl mono-substituted, di-substituted or unsubstituted amino;

wherein R³¹ is independently hydrogen, cyano, nitro, hydroxy, C₁-C₂₀ alkyl, C₂-C₂₀ alkenyl, C₂-C₂₀ alkynyl, C₃-C₁₂ cycloalkyl, C₁-C₉ heterocycloalkyl, C₆-C₁₂ aryl, or C₁-C₉ heteroaryl;

wherein R³² is independently C₁-C₂₀ alkyl, C₂-C₂₀ alkenyl, C₂-C₂₀ alkynyl, or substituted or unsubstituted C₁-C₂₀ silyl; wherein the substituent in the “substituted or unsubstituted C₁-C₂₀ silyl” is C₂-C₂₀ alkenyl or C₂-C₂₀ alkynyl;

wherein each of R^(33a) and R^(33b) is independently hydrogen, hydroxy, cyano, nitro, halogen, C₁-C₂₀ alkyl, C₂-C₂₀ alkenyl, C₂-C₂₀ alkynyl, C₁-C₂₀ alkoxy, C₃-C₁₂ cycloalkyl, C₁-C₉ heterocycloalkyl, C₆-C₁₂ aryl, or C₁-C₉ heteroaryl;

R^(6a), R^(6b), R^(6c) and R^(6d) are independently hydrogen, hydroxy, C₁-C₂₀ alkyl, C₃-C₁₂ cycloalkyl or C₃-C₁₂ heterocycloalkyl, but R^(6a) and R^(6b) are not hydroxy at the same time, R^(6c) and R^(6d) are not hydroxyl at the same time; or R^(6a) and R^(6b) together with the carbon atom attached form carbonyl; or R^(6c) and R^(6d) together with the carbon atom attached form carbonyl;

the number of the heteroatom in the “heterocycloalkyl”, “heteroaryl” is 1 to 7 selected from the group consisting of oxygen, nitrogen, phosphorus, sulfur, selenium, boron and silicon;

and, the compound I is not selected from the group consisting of

Preferably, the C₂-C₂₀ alkenyl contained in the term “substituted or unsubstituted C₂-C₂₀ alkenyl” and the term “C₂-C₂₀ alkenyl” is independently C₂-C₁₂ alkenyl; more preferably, the C₂-C₂₀ alkenyl contained in the term “substituted or unsubstituted C₂-C₂₀ alkenyl” is independently C₂-C₆ alkenyl.

Preferably, the C₂-C₂₀ alkynyl contained in the term “substituted or unsubstituted C₂-C₂₀ alkynyl” and the term “C₂-C₂₀ alkynyl” is independently C₂-C₁₂ alkynyl; more preferably, C₂-C₂₀ alkynyl contained in the term “substituted or unsubstituted C₂-C₂₀ alkynyl” is independently C₂-C₆ alkynyl.

Preferably, the C₁-C₂₀ alkyl contained in the term “substituted or unsubstituted C₁-C₂₀ alkyl”, “C₁-C₂₀ alkyl” and “C₁-C₂₀ alkyl mono-substituted, di-substituted or unsubstituted amino” is independently C₁-C₁₂ alkyl; more preferably, the C₂-C₂₀ alkyl contained in the term “substituted or unsubstituted C₁-C₂₀ alkyl” is independently C₁-C₆ alkyl; most preferably, the C₂-C₂₀ alkyl contained in the term “substituted or unsubstituted C₁-C₂₀ alkyl” is independently C₁-C₃ alkyl.

Preferably, the C₁-C₂₀ alkylthio contained in the term “substituted or unsubstituted C₁-C₂₀ alkylthio” and “C₁-C₂₀ alkylthio” is independently C₁-C₁₂ alkylthio; more preferably, the C₂-C₂₀ alkylthio contained in the term “substituted or unsubstituted C₁-C₂₀ alkylthio” is independently C₁-C₆ alkylthio; most preferably, the C₂-C₂₀ alkylthio contained in the term “substituted or unsubstituted C₁-C₂₀ alkylthio” is independently C₁-C₃ alkylthio.

Preferably, the C₁-C₂₀ alkoxy contained in the term “substituted or unsubstituted C₁-C₂₀ alkoxy” and the term “C₁-C₂₀ alkoxy” is independently C₁-C₁₂ alkoxy; more preferably, the C₁-C₂₀ alkoxy contained in the term “substituted or unsubstituted C₁-C₂₀ alkoxy” is independently C₁-C₆ alkoxy; most preferably, the C₁-C₂₀ alkoxy contained in the term “substituted or unsubstituted C₁-C₂₀ alkoxy” is independently C₁-C₃ alkoxy.

Preferably, the “C₁-C₂₀ silyl” is independently “C₁-C₁₂ silyl”; more preferably, the “C₁-C₂₀ silyl” is independently “C₁-C₆ silyl”; most preferably, the “C₁-C₂₀ silyl” is independently “C₁-C₃ silyl”.

Preferably, the C₃-C₁₂ cycloalkyl contained in the term “substituted or unsubstituted C₃-C₁₂ cycloalkyl” is independently C₃-C₈ cycloalkyl; more preferably, the C₃-C₁₂ cycloalkyl contained in the term “substituted or unsubstituted C₃-C₁₂ cycloalkyl” is independently C₃-C₆ cycloalkyl.

Preferably, the C₁-C₉ heterocycloalkyl contained in the term “substituted or unsubstituted C₁-C₉ heterocycloalkyl” is independently C₃-C₈ heterocycloalkyl having 1 to 4 (e.g. 1 to 3) heteroatom selected from the group consisting of boron, silicon, oxygen, sulfur, selenium, nitrogen and phosphorus; more preferably, the C₁-C₉ heterocycloalkyl contained in the term “substituted or unsubstituted C₁-C₉ heterocycloalkyl” is independently C₃-C₅ heterocycloalkyl having 1 to 4 (e.g. 1 to 3) heteroatom selected from the group consisting of boron, silicon, oxygen, sulfur, selenium, nitrogen and phosphorus.

Preferably, the C₁-C₉ heteroaryl contained in the term “substituted or unsubstituted C₁-C₉ heteroaryl” is independently C₁-C₆ heteroaryl; more preferably, the C₁-C₉ heteroaryl contained in the term “substituted or unsubstituted C₁-C₉ heteroaryl” is independently C₁-C₆ heteroaryl having 1 to 4 heteroatom selected from the group consisting of boron, silicon, oxygen, sulfur, selenium, nitrogen and phosphorus.

Preferably, the C₃-C₁₂ aryl contained in the term “substituted or unsubstituted C₃-C₁₂ aryl” is independently C₆-C₁₀ aryl.

In some embodiments, the definitions are preferably as follows:

In the compound I, preferably, p is 1, q is 1, m is 0 or 1, n is 0.

In the compound I, preferably, X is N.

In the compound I, preferably, R¹ is halogen or trifluoromethyl.

In the compound I, preferably, R² is C₁-C₆ alkyl.

In the compound I, preferably, R³ is substituted or unsubstituted C₁-C₆ alkyl, or substituted or unsubstituted C₃-C₈ cycloalkyl.

In the compound I, preferably, R⁴ is halogen.

In the compound I, preferably, R⁵ is hydrogen, substituted or unsubstituted C₁-C₆ alkyl,

In the compound I, preferably, R⁸ is independently substituted or unsubstituted C₁-C₆ alkyl, or C₁-C₆ alkoxy.

In the compound I, preferably, R⁹ is vinyl, substituted or unsubstituted C₁-C₆ alkyl, substituted or unsubstituted C₃-C₈ cycloalkyl, substituted or unsubstituted heterocycloalkyl having 3 to 8 carbon atoms and 1 to 4 heteroatom selected from the group consisting of oxygen, sulfur and nitrogen, substituted or unsubstituted C₁-C₆ alkoxy or

In the compound I, preferably, R^(10a) and R^(10b) are independently hydrogen, or substituted or unsubstituted C₁-C₆ alkyl.

In the compound I, preferably, the “substituted” in the “substituted or unsubstituted C₁-C₂₀ alkyl”, “substituted or unsubstituted C₃-C₁₂ cycloalkyl”, “substituted or unsubstituted C₁-C₉ heterocycloalkyl” and “substituted or unsubstituted C₁-C₂₀ alkoxy” in the definitions of R⁵, R⁸, R⁹, R^(10a) and R^(10b) refers to be independently substituted with the substituent selected from the group consisting of

hydroxy, halogen, C₁-C₃ alkoxy, “substituted or unsubstituted C₁-C₉ saturated or unsaturated heterocycloalkyl” and

when there are more than one substituents, the substituents are the same or different.

In the compound I, preferably, R^(11a) and R^(11b) are independently hydrogen, or C₁-C₆ alkyl.

In the compound I, preferably, R¹² is hydroxy, C₁-C₆ alkoxy or

In the compound I, preferably, R^(27a) and R^(27b) are independently C₁-C₆ alkyl.

In the compound I, preferably, R^(6a), R^(6b), R^(6c) and R^(6d) are independently hydrogen; or R^(6c) and R^(6d) together with the carbon atom attached form carbonyl.

In some embodiments, the definitions are preferably as follows:

p is preferably 1.

q is preferably 1.

n is preferably 0.

m is preferably 1.

X is preferably N.

Y is preferably CH.

N atom in the cycle

is preferably oxidized or non-oxidized;

R^(6a), R^(6b), R^(6c) and R^(6d) are preferably hydrogen.

R¹ is preferably halogen (e.g. fluorine).

R² is preferably C₁-C₆ alkyl.

R³ is preferably substituted or unsubstituted C₁-C₂₀ alkyl (e.g. isopropyl or tert-butyl), or substituted or unsubstituted C₃-C₁₂ cycloalkyl(e.g. 1-methylcyclopropyl or cyclopropyl).

R⁴ is preferably halogen (e.g. fluorine).

R⁵ is preferably substituted C₁-C₂₀ alkyl (e.g. substituted ethyl; e.g. substitute disopropyl).

The “substituted” in the “substituted C₁-C₂₀ alkyl” in the definition of R⁵ refers to preferably be substituted with the substituent selected from the group consisting of substituted or unsubstituted C₁-C₉ saturated or unsaturated heterocycloalkyl,

when there are more than one substituents, the substituents are the same or different; N atom in

is oxidized or non-oxidized.

Preferably, R^(11a) and R^(11b) are independently hydrogen, substituted or unsubstituted C₁-C₂₀ alkyl(e.g. substituted or unsubstituted C₁-C₆ alkyl, preferably substituted or unsubstituted C₁, C₂ or C₃ alkyl; e.g. —CF₃; e.g. hydroxyethyl, 2-dimethylaminoethyl), or

R^(11a) and R^(11b) together with the N atom attached form

wherein t and u are independently 1, 2 or 3, R^(c) is C(R^(cs))², NR^(cs), O, S, Si(R^(cs))₂ or Se, each R^(cs) is independently H, C₁-C₆ alkyl (e.g. methyl, ethyl, propyl or isopropyl), or B(OH)₂; or in

R^(11a) and R^(11b) together with the N atom attached form

wherein v and w are independently 1, 2 or 3, R^(ch) is CH or PH₂, R^(cx) is CH₂, O, S or ═N—OH, R^(cx) is preferably attached to R^(ch)

R^(hc) is H or C₁-C₆ alkyl (e.g. methyl, ethyl or propyl), preferably when R^(ch) is P, R^(ch) is C₁-C₆ alkyl.

Preferably, R¹⁵ is C₁-C₆ alkyl.

Preferably, the “substituted” in the term “substituted or unsubstituted C₁-C₂₀ alkyl” in the definitions of R^(11a) and R^(11b) refers to be independently substituted with the substituent selected from the group consisting of hydroxy, C₁-C₆ silyl,

when there are more than one substituents, the substituents are the same or different.

R^(24a) is preferably hydrogen.

R^(24b) is preferably C₁-C₆ alkyl.

Preferably, R^(25a) and R^(25b) are independently hydroxyl or C₁-C₆ alkoxy (e.g. ethoxy).

Preferably, R^(27a) and R^(27b) are independently C₁-C₆ alkyl.

Preferably, the compound I in the present invention is selected from the group consisting of

The nitrogen-containing fused heterocyclic compounds involved in the present invention may exhibit tautomerism, structural isomerism and stereoisomerism. The present invention includes any tautomeric or structural isomeric or stereoisomeric forms thereof and mixtures thereof, which have high selectivity and high inhibitory activity on CDK4 and CDK6, at the same time, they have better inhibitory activity against breast cancer cells, which is not limited to any of the isomers or the mixture thereof.

The nitrogen-containing fused heterocyclic compounds in the present invention can be prepared by various methods well-known to those skilled in the field of organic synthesis and medicinal chemistry, or prepared by the methods described below combined with the synthetic methods known in the field of organic chemistry or transformational methods thereof understood by those skilled in the art.

The preparation method for the nitrogen-containing fused heterocyclic compounds can start from readily available starting materials using the following general methods and processes to prepare the compound in the present invention. It will be understood that where typical or preferable process operating conditions (e.g. reaction temperature, time, mole ratio of reactants, solvent, pressure, etc.) are given; other process operating conditions can also be used, unless otherwise specified.

Optimum reaction conditions may vary with the specific reactant or solvent used, but these conditions can be determined by those skilled in the art by routine optimization procedures.

The preparation method for the nitrogen-containing fused heterocyclic compounds described herein can be monitored by any suitable method known in the art. For example, the product formation can be monitored by nuclear magnetic resonance, infrared spectroscopy, spectrophotometry or mass spectrometry, HPLC, or thin layer chromatography.

The preparation method for the nitrogen-containing fused heterocyclic compound may involve protection and deprotection of multiple chemical groups. The need for protection and deprotection, as well as the choice of the appropriate protecting group, can be easily determined by those skilled in the art. The chemical process of the protecting group can refer to Greene et al., Protective Groups in Organic Synthesis, Second Edition, Wiley & Sons, 1991, which is incorporated herein by reference in its entirety.

The preparation method described herein can be carried out in a suitable solvent, which can be readily selectable by those skilled in the art of organic synthesis.

A suitable solvent does not substantially react with the starting materials, intermediates or products at the temperature at which the described reaction is carried out. The temperature at which the reaction is carried out may vary from the solvent's freezing point to the solvent's boiling temperature. The reaction can be carried out in a solvent or a mixture of multiple solvents. Depending on a specific reaction step, the solvent suitable for the specific reaction step can be selected.

On this basis, particularly preferably (but not limited to reagents and solvents in the reaction condition), the present invention further provides a preparation method for the nitrogen-containing fused heterocyclic compound, which is any one of the following methods:

Method 1:

when R⁵ is hydrogen, the preparation method for the compound represented by formula I comprises conducting Suzuki coupling reaction to give 1C, followed by conducting Buchwald coupling with the compound represented by formula 1C and

to remove the protecting group and give the compound represented by formula I; the PG in the compound represented by formula 1D is a protecting group;

wherein X, Y, m, n, p, q, R¹, R², R³, R⁴, R^(6a), R^(6b), R^(6c) and R^(6d) are defined as above;

the PG in the compound represented by formula 1D can be various conventional amino protecting groups in the art, preferably Boc, for the purpose of making certain reactive groups (e.g. amino group) not participate in the reaction when

reacts with compound 1C;

the condition of the deprotection reaction can be conventional removal conditions for various protecting groups in the art, e.g. conditions of hydrolysis reaction, conditions of amine hydrolysis reaction, conditions of hydrogenation reaction, and the like;

after the completion of deprotection reaction, preferably further comprising post-treatment; the method and condition of the post-treatment can be conventional methods and conditions for post-reaction treatment in the art, preferably comprising washing the reaction system, drying, filtering, evaporating to dryness; then conducting column chromatography; or, the reaction system is distilled to remove the solvent, washed, filtered; or, the reaction system is distilled to remove the solvent, subjected to thin layer chromatography;

the condition of the substitution reaction or the transition metal catalyzed coupling reaction can be various conventional conditions of such reactions in the art; the substitution reaction may be carried out by heating or pressurizing or acid-base catalyzing etc.;

wherein, the condition of each reaction step in the reaction route can be carried out according to conventional conditions of such reactions in the art;

Method 2:

when R⁵ is hydrogen, a preparation method for the compound represented by formula I comprises subjecting the 1C obtained according to method 1 to ammonolysis reaction, then reacting with

to give 2A, followed by removing the protecting group to give the compound represented by formula I;

wherein X, Y, m, n, p, q, R¹, R², R³, R⁴, R^(6a), R^(6b), R^(6c) and R^(6d) are defined as above;

Method 3:

when R⁵ is

and R¹¹ is substituted or unsubstituted C₁-C₆ alkyl, or substituted or unsubstituted C₃-C₈ cycloalkyl, the preparation method for the compound represented by formula I comprises that in organic solvent (e.g. selected from the group consisting of 1,4-dioxane, dichloromethane and DMF), in the presence of a condensing agent (e.g. HOBt and EDCI), conducting condensation reaction with compound 3A and

to give compound I; the condition of the condensation reaction can be conventional conditions in the art;

X, Y, m, n, p, q, R¹, R², R³, R⁴, R, R^(6a), R^(6b), R^(6c) and R^(6d) are defined as above;

Method 4:

when R⁵ is

the preparation method for the compound represented by formula I comprises that in organic solvent (e.g. tetrahydrofuran), in the presence of the base (e.g. pyridine), reacting compound 4A with

to give compound 4B, followed by conducting hydrolysis reaction to give compound I; the conditions of the amide reaction and the hydrolysis reaction can be conventional conditions in the art;

X, Y, m, n, p, q, R¹, R², R³, R⁴, R^(6a), R^(6b), R^(6c), R^(6d) and R⁸ are defined as above;

Method 5:

when R⁵ is

R¹¹ is substituted C₁-C₆ alkyl or C₃-C₈ cycloalkyl, and the substituent is

when R^(13b) is H, the preparation method for the compound represented by formula I comprises that in organic solvent (e.g. selected from the group consisting of 1,4-dioxane, dichloromethane and DMF), in the presence of condensing agent (e.g. HOBt and EDCI), reacting compound 5A with

to give compound 5B, then removing the protecting group of the amino group to give compound I; the condition of the amide reaction and the deprotection reaction can be conventional conditions in the art; R^(13a)NH—(CH₂)_(n1)—C(═O)— is R⁵; —(CH₂)_(n1)— is the substituted C₁-C₆ alkyl or C₃-C₈ cycloalkyl;

X, Y, m, n, p, q, R¹, R², R³, R⁴, n₁, R^(6a), R^(6b), R^(6c), R^(6d) and R^(13a) are defined as above;

PG in the compound represented by formula 5B can be various conventional amino protecting groups in the art, preferably Boc, for the purpose of making certain reactive groups (e.g. amino group) not participate in the reaction when

reacts with compound 5A;

Method 6:

when R⁵ is

R¹¹ is substituted C₁-C₆ alkyl or C₃-C₈ cycloalkyl, the substituent is

and neither R^(13a) or R^(13b) is H, the preparation method for the compound represented by formula I comprises that in organic solvent (e.g. selected from the group consisting of 1,4-dioxane, dichloromethane and DMF), in the presence of condensing agent (e.g. HOBt and EDCI), reacting compound 6A with

to give compound 6B, then removing the protecting group of the amino group and conducting reductive amination to give compound I; the conditions of the amide reaction, the deprotection reaction and the reductive amination can be conventional conditions in the art;

is R⁵;

X Y, m, n, p R¹, R², R³, R⁴, R^(6a), R^(6b), R^(6c), R^(6d), R^(13a) and R^(13b) are defined as above;

PG in the compound represented by formula 6B can be various conventional amino protecting groups in the art, preferably Boc, for the purpose of making certain reactive groups (e.g. amino group) not participate in the reaction when

reacts with compound 6A;

Method 7:

when R⁵ is

R¹¹ is substituted C₁-C₆ alkyl or C₃-C₈ cycloalkyl, and the substituent is

R^(13b) and R^(13b) are the same, the preparation method for the compound represented by formula I comprises that in organic solvent (e.g. selected from the group consisting of 1,4-dioxane, dichloromethane and DMF), in the presence of condensing agent (e.g. HOBt and EDCI), reacting compound 7A with

to give compound 7B, then removing the protecting group of the amino group and conducting reductive amination to give compound I; the conditions of the amide reaction, the deprotection reaction and the reductive amination can be conventional conditions in the art;

is R⁵;

X, Y, m, n, p, q, R¹, R², R³, R⁴, R^(6a), R^(6b), R^(6c), R^(6d) and R^(13a) are defined as above;

PG in the compound represented by formula 7B can be various conventional amino protecting groups in the art, preferably Boc, for the purpose of making certain reactive groups (e.g. amino group) not participate in the reaction when

reacts with compound I;

Method 8:

when R⁵ is substituted or unsubstituted C₁-C₆ alkyl, substituted or unsubstituted C₃-C₈ cycloalkyl

and substituted or unsubstituted C₃-C₈ heterocycloalkyl, the preparation method for the compound represented by formula I comprises that in organic solvent (e.g. DMF and/or 1,4-dioxane), in the presence of base (e.g. N,N-diisopropylethylamine), conducting substitution reaction with compound 8A and R⁵—X to give compound I; the conditions of the substitution reaction can be conventional conditions in the art; X is halogen (e.g. bromine) or CH₃SO₃—;

X, Y, m, n, p, q, R¹, R², R³, R⁴, R⁵, R^(6a), R^(6b), R^(6c) and R^(6d) are defined as above;

Method 9:

when R⁵ is substituted or unsubstituted C₁-C₆ alkyl, substituted or unsubstituted C₃-C₈ cycloalkyl

or substituted or unsubstituted C₃-C₈ heterocycloalkyl, the preparation method for the compound represented by formula I comprises that in organic solvent (e.g. selected from the group consisting of 1,2-dichloroethane, methanol and dioxane), in the presence of reductive agent (e.g. sodium triacetoxyborohydride and/or sodium cyanoborohydride), conducting reductive amination reaction with compound 9A and R^(5a)—CHO to give compound I; the condition of the reductive amination reaction can be conventional conditions in the art; R^(5a) CH₂— is R⁵;

X, Y, m, n, p, q, R¹, R², R³, R⁴, R⁵, R^(6a), R^(6b), R^(6c) and R^(6d) are defined as above;

Method 10:

when R⁵ is substituted C₁-C₆ alkyl, substituted C₃-C₈ cycloalkyl, or, substituted C₃-C₈ heterocycloalkyl, the substituent is

R¹⁶ is hydroxyl, the preparation method for the compound represented by formula I comprises that in organic solvent (e.g. selected from the group consisting of DMF, dichloromethane and 1,4-dioxane), in the presence of base (e.g. N,N-diisopropylethylamine), conducting substitution reaction with compound 10A and R^(5b)—OC(═O)—(CH₂)_(n2)—X to give compound I; the condition of the substitution reaction and the hydrolysis reaction can be conventional conditions in the art; X is halogen (e.g. bromine) or CH₃SO₃—; R^(5b) is C₁-C₆ alkyl(e.g. methyl); —(CH₂)_(n2) is the substituted C₁-C₆ alkyl(can be branched alkyl or straight alkyl) in R⁵, substituted C₃-C₈ cycloalkyl, or, substituted C₃-C₈ heterocycloalkyl;

is R⁵;

X, Y, Z, m, n, p, q, R¹, R², R³, R⁴, n₂, R^(6a), R^(6b), R⁶ and R^(6d) are defined as above;

Method 11:

when R⁵ is substituted C₁-C₆ alkyl, substituted C₃-C₈ cycloalkyl, the substituent is

(wherein at least one of R^(13a) and R^(13b) is hydrogen) or hydroxyl, or, unsubstituted C₃-C₈ heterocycloalkyl, the preparation method for the compound represented by formula I comprises that in organic solvent (e.g. selected from the group consisting of 1,2-dichloroethane, methanol and/or dioxane), in the presence of reductive agent (e.g. sodium triacetoxyborohydride and/or sodium cyanoborohydride), conducting reductive amination reaction with compound 11A and PG-R^(5c)—CHO, then removing the protecting group to give compound I; the conditions of the reductive amination reaction and the deprotection reaction can be conventional conditions in the art; R^(5c)CH₂— is R⁵;

X, Y, Z, m, n, p, q, R¹, R², R³, R⁴, n₂, R^(6a), R^(6b), R⁶ and R^(6d) are defined as above;

PG in the compound represented by formula 11B can be various conventional amino protecting groups in the art, preferably Boc, for the purpose of making certain reactive groups (e.g. amino group) not participate in the reaction when PG-R^(5c)—CHO reacts with compound 11A;

Method 12;

when R⁵ is

R¹¹ is

and R^(12a) and R^(12b) are independently substituted or unsubstituted C₁-C₂₀ alkyl, the preparation method for the compound represented by formula I comprises that in organic solvent, in the presence of triphosgene, conducting condensation reaction with compound 11A and substituted or unsubstituted amino to give compound I; R^(12a)NR^(12b)—C(═O)— is R⁵;

X, Y, Z, m, n, p, q, R¹, R², R³, R⁴, n₂, R^(6a), R^(6b), R⁶ and R^(6d) are defined as above;

Method 13:

when R⁵ is substituted C₁-C₂₀ alkyl, or, substituted C₁-C₉ heterocycloalkyl, the substituent is

C₁-C₂₀ alkyl, C₂-C₂₀ alkenyl, C₂-C₂₀ alkynyl, substituted C₁-C₉ heterocycloalkyl, or, C₃-C₁₂ cycloalkyl, and R^(3a) and R^(13b) are independently hydrogen, C₁-C₂₀ alkyl, C₂-C₂₀ alkenyl, C₂-C₂₀ alkynyl, C₃-C₁₂ cycloalkyl, borono, phospho, guanidino, thioureido,

the preparation method for the compound represented by formula I comprises that in organic solvent, in the presence of reductive agent, conducting reductive amination reaction with compound 13A and PG-NR^(13a)—(CH₂)n-CHO, then removing the protecting group, followed by substitution reaction to give compound I; R^(13a)NR^(13b)—(CH₂)n- is R⁵; LG in the formula of the compound is a leaving group;

X, Y, m, n, p, q, R¹, R², R³, R⁴, R⁵, R^(6a), R^(6b), R^(6c) and R^(6d) are defined as above;

Method 14:

when R⁵ is substituted C₁-C₂₀ alkyl, or, substituted C₁-C₉ heterocycloalkyl, the substituent is

C₁-C₂₀ alkyl, C₂-C₂₀ alkenyl, C₂-C₂₀ alkynyl, substituted C₁-C₉ heterocycloalkyl, or, C₃-C₁₂ cycloalkyl, and R^(14a), R^(14b) and R^(14c) are independently hydrogen, C₁-C₂₀ alkyl, C₂-C₂₀ alkenyl, C₂-C₂₀ alkynyl, C₃-C₁₂ cycloalkyl, borono, phospho, guanidino, thioureido,

the preparation method for the compound represented by formula I comprises that in organic solvent, in the presence of reductive agent, conducting reductive amination reaction with compound 14A and PG-NR^(14a)—(CH₂)n-CHO, then removing the protecting group, followed by substitution reaction to give compound I; R^(14b)NR^(14c)—(CH₂)n-R^(14a) is R⁵; LG in the formula of compound is a leaving group;

X, Y, m, n, p, q, R¹, R², R³, R⁴, R⁵, R^(6a), R^(6b), R^(6c) and R^(6d) are defined as above;

Method 15:

when R⁵ is substituted C₁-C₂₀ alkyl, or, substituted C₁-C₉ heterocycloalkyl, the substituent is

C₁-C₂₀ alkyl, C₂-C₂₀ alkenyl, C₂-C₂₀ alkynyl, substituted C₁-C₉ heterocycloalkyl, or, C₃-C₁₂ cycloalkyl, and R^(13a) and R^(13b) are independently hydrogen, C₁-C₂₀ alkyl, C₂-C₂₀ alkenyl, C₂-C₂₀ alkynyl, C₃-C₁₂ cycloalkyl, borono, phospho, guanidino, thioureido,

the preparation method for the compound represented by formula I comprises that in organic solvent, in the presence of reductive agent, conducting substitution reaction with compound 15A and LG-(CH₂)n-CN, followed by addition reaction to give compound I; LG in the formula compound is a leaving group;

X, Y, m, n, p, q, R¹, R², R³, R⁴, R⁵, R^(6a), R^(6b), R^(6c) and R^(6d) are defined as above;

after the completing of the preparation method for the nitrogen-containing fused heterocyclic compound, the post-treatment is preferably conducted; the methods and conditions of the post-treatment can be conventional methods and conditions for the post-reaction treatment in the art, preferably comprising washing the reaction system, drying, filtering, evaporating to dryness, then being subjected to column chromatography; or, the reaction system is distilled to remove the solvent, washed, and filtered; or the reaction system is distilled to remove the solvent, subjected to thin layer chromatography.

The present invention also provides a compound II, which is selected from the group consisting of

wherein m, n, p, q, X, Y, R¹, R², R³, R⁴, R⁵, R^(5a), R^(5b), R^(5c), R^(6a), R^(6b), R^(6c), R^(6d), R⁸, R^(13a) and PG are defined as above.

Preferably, the compound II is selected from the group consisting of

The present invention also provides a use of the nitrogen-containing fused heterocyclic compound, the pharmaceutically acceptable salt, the enantiomer, the diastereomer, the tautomer, the solvate, the metabolite or the drug precursor thereof in manufacturing medicament, the medicament is used for the prevention or the treatment of disorder associated with abnormal cell cycle regulation; the “disorder associated with abnormal cell cycle regulation” is preferably “disorder associated with abnormality of cyclin-dependent kinase (preferably CDK4 and/or CDK6)”, more preferably tumor, most preferably malignant tumor (e.g. breast cancer).

The present invention also provides a use of the nitrogen-containing fused heterocyclic compound, the pharmaceutically acceptable salt, the enantiomer, the diastereomer, the tautomer, the solvate, the metabolite or the drug precursor thereof in manufacturing an inhibitor of cyclin-dependent kinase (preferably CDK4 and/or CDK6).

The present invention also provides a use of the nitrogen-containing fused heterocyclic compound, the pharmaceutically acceptable salt, the enantiomer, the diastereomer, the tautomer, the solvate, the metabolite or the drug precursor thereof in manufacturing medicament having tumor cell inhibitory activity; the tumor cell is preferably cancer cell; the cancer cell is preferably breast cancer cell; the breast cancer cell is preferably selected from the group consisting of breast cancer cell MCF-7, T-47D and ZR-75-1.

The present invention also provides a composition, which comprises the nitrogen-containing fused heterocyclic compound, the pharmaceutically acceptable salt, the enantiomer, the diastereomer, the tautomer, the solvate, the metabolite or the drug precursor thereof, and at least a pharmaceutical excipient.

The dose of the nitrogen-containing fused heterocyclic compound, the pharmaceutically acceptable salt, the enantiomer, the diastereomer, the tautomer, the solvate, the metabolite or the drug precursor thereof can be a therapeutically effective dose.

The select of the pharmaceutical excipient varies depending on the route of administration and the characteristic of action, can generally be fillers, diluents, adhesives, wetting agents, disintegrants, lubricants, emulsifiers, suspending agents etc. that are conventional in the art.

The route of administration of the pharmaceutical composition can be oral administration, injection (intravenous, intramuscular, subcutaneous and intracoronary), sublingual administration, buccal administration, rectal administration, transurethral administration, transvaginal administration, nasal administration, inhaled administration or topical administration, preferably oral administration.

In the present invention, unless otherwise specified, the following terms in the description and the claims of the invention have the following meanings:

The term “halogen” is preferably fluorine, chlorine, bromine, iodine, more preferably fluorine.

The term “C₁-C₂₀ alkyl” refers to a straight chain or branched saturated hydrocarbon group containing 1 to 20 carbon atoms. Alkyl can be optionally substituted with one or more than one substituent described in the present invention. Examples of alkyl include but not limited to methyl (Me, —CH₃), ethyl (Et, —CH₂CH₃), n-propyl (n-Pr, —CH₂CH₂CH₃), isopropyl (i-Pr, —CH(CH₃)₂), n-butyl(n-Bu, —CH₂CH₂CH₂CH₃), 2-methylpropyl or isobutyl (i-Bu, —CH₂CH(CH₃)₂), 1-methylpropyl or sec-butyl (s-Bu, —CH(CH₃)CH₂CH₃), tert-butyl (t-Bu, —C(CH₃)₃), n-pentyl (—CH₂CH₂CH₂CH₂CH₃), 2-pentyl (—CH(CH₃)CH₂CH₂CH₃), 3-pentyl (—CH(CH₂CH₃)₂), 2-methyl-2-butyl(-C(CH₃)₂CH₂CH₃), 3-methyl-2-butyl(-CH(CH₃)CH(CH₃)₂), 3-methyl-1-butyl(-CH₂CH₂CH(CH₃)₂), 2-methyl-1-butyl(-CH₂CH(CH₃)CH₂CH₃), n-hexyl (—CH₂CH₂CH₂CH₂CH₂CH₃), 4-methylpentyl(-CH₂CH₂CH₂CH(CH₃)CH₃), 3-methylpentyl (—CH₂CH₂CH(CH₃)CH₂CH₃), 2-methylpentyl (—CH₂CH(CH₃)CH₂CH₂CH₃), 2-hexyl (—CH(CH₃)CH₂CH₂CH₂CH₃), 3-hexyl (—CH(CH₂CH₃)(CH₂CH₂CH₃), 3,3-dimethylbutyl (—CH₂CH₂CH₂(CH₃)₂CH₃), 2,2-dimethylbutyl (—CH₂C(CH₃)₂CH₂CH₃), 2-methyl-2-pentyl (—C(CH₃)₂CH₂CH₂CH₃), 3-methyl-2-pentyl (—CH(CH₃)CH(CH₃)CH₂CH₃), 4-methyl-2-pentyl (—CH(CH₃)CH₂CH(CH₃)₂), 3-methyl-3-pentyl (—C(CH₃)(CH₂CH₃)₂), 2-methyl-3-pentyl (—CH(CH₂CH₃)CH(CH₃)₂), 2,3-dimethyl-2-butyl(-C(CH₃)₂CH(CH₃)₂), 3,3-dimethyl-2-butyl(-CH(CH₃)C(CH₃)₃), n-heptyl, n-octyl etc.

In the present invention, the term “alkenyl” refers to a straight, branched, or cyclic non-aromatic hydrocarbon group containing the specified number of carbon atoms and at least one carbon-carbon double bond. Alkenyl is preferably containing one carbon-carbon double bond, and up to four non-aromatic carbon-carbon double bonds can be present. Thus, “C₂₋₁₂ alkenyl” refers to an alkenyl group having 2 to 12 carbon atoms. “C₂₋₆ alkenyl” refers to an alkenyl group having 2 to 6 carbon atoms, including vinyl, propenyl, butenyl, 2-methylbutenyl and cyclohexenyl. The straight chain, branched chain or cyclic portion of alkenyl can contain double bond, and if it is substituted alkenyl, the alkenyl can be substituted.

The term “alkynyl” refers to a straight, branched, or cyclic hydrocarbon group containing the specified number of carbon atoms and at least one carbon-carbon triple bond. Up to three carbon-carbon triple bonds can be present. Thus, “C₂₋₁₂ alkynyl” refers to an alkynyl group having 2 to 12 carbon atoms. “C₂₋₆ alkynyl” refers to an alkynyl group having 2 to 6 carbon atoms, including but not limited to ethynyl, propynyl, butynyl, and 3-methylbutynyl.

The term “C₁-C₂ alkoxy” refers to C₁-C₂₀ alkyl connected by the oxygen atom; the C₁-C₂₀ alkyl is defined as above.

The term “C₁-C₂₀ alkylthio” refers to C₁-C₂₀ alkyl connected by the sulfur atom; the C₁-C₂₀ alkyl is defined as above.

The term “C₁-C₂₀ silyl” refers to C₁-C₂₀ alkyl connected by the silicon atom; the C₁-C₂₀ alkyl is defined as above.

The term “C₃-C₁₂ cycloalkyl” refers to a cyclic hydrocarbon group that contains 3 to 12 ring-forming carbon atoms, can be saturated or partially unsaturated (contain 1 or 2 double bonds, but none of the rings have a fully conjugated 7 electron system), and does not contain heteroatom; including a monocycle containing 3 to 12 carbon atoms or a bicycle or tricycle containing 7 to 12 carbon atoms (including spiro ring, bridged ring and fused ring); wherein one or more than one hydrogen atoms in the ring can be independently optionally substituted with one or more than one substituent described herein, and the carbon atoms can be oxidized. A bicyclic carbon ring containing 7 to 12 atoms can be bicyclic[4, 5], [5,5], [5,6] or [6,6] system, and a bicyclic carbon ring containing 9 or 10 atoms can be bicyclic[5,6] or [6,6] system. Suitable cycloalkyl group include, but is not limited to, cycloalkyl, cycloalkenyl and cycloalkynyl, e.g. cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 1-cyclopentyl-1-enyl, 1-cyclopentyl-2-enyl, 1-cyclopentyl-3-enyl, cyclohexyl, 1-cyclohexyl-1-enyl, 1-cyclohexyl-2-alkenyl, 1-cyclohexyl-3-enyl, cyclohexadienyl, cycloheptyl, cyclooctyl, cyclononyl, cyclononyl, cycloundecyl, cyclododecyl, adamantyl etc. Depending on the structure, cycloalkyl can be a monovalent or divalent group, i.e. cycloalkylene.

The term “heterocycloalkyl” refers to a 3 to 10 membered monocyclic or polycyclic group (including spiro ring, bridged ring and fused ring) containing 1 to 6 heteroatoms (selected from the group consisting of N, S, B, P, Si, O and Se), wherein each ring can contain one or more than one double bonds, but none of the rings has a fully conjugated 7 electron system; heteroatom can be substituted or unsubstituted, and N atom can be quaternized. Heterocyclic system can be attached to the main structure via any heteroatom or carbon atom to form a stable compound. One or more than one hydrogen atoms on the ring are independently optionally substituted with one or more than one substituent described herein. For example, 3 to 7 membered monocycle (1 to 6 carbon atoms and 1 to 3 heteroatoms selected from the group consisting of N, O, P, B, Si, S and Se, wherein N, S, B, P or Se is optionally substituted with one or more than one oxygen atoms to form groups like NO, NO₂, BOH, SO, SO₂, PO, PO₂ and SeO, meanwhile —CH₂— group can be optionally replaced with —C(═O)—, —C(═S)— or —C(═N)—; —SH₂— group can be optionally replaced with —S(═O)—, —S(═O)₂—, —S(═N)— or —S(═N)₂—; when the ring is a 3 membered ring, there is only one heteroatom), or a bicyclic ring formed by 7 to 10 atoms (4 to 9 carbons atoms and 1 to 3 heteroatoms selected from the group consisting of N, O, P, B, Si and S, wherein N, S, B or P is optionally substituted with one or more than one oxygen atom to form groups like NO, NO₂, BOH, SO, SO₂, PO, PO₂, SeO, meanwhile —CH₂— group can be optionally replaced with —C(═O)—. Depending on the structure, the heterocyclic group can be a monovalent or a divalent group, i.e. heterocyclylene. In some embodiments, N atom in the nitrogen-containing heterocycle can be oxidized to form nitrogen oxide.

The term “heteroaryl” refers to a 3 to 10 membered monocyclic or polycyclic aromatic system containing 1 to 6 heteroatoms (selected from the group consisting of N, S, B, P, Si, O and Se, wherein bicyclic heteroaromatic ring, tricyclic heteroaromatic ring or tetracyclic heteroaromatic ring system forms the ring in a fused form, and wherein N, S, B, P or Se is optionally substituted with one or more than one oxygen atom to form groups like NO, NO₂, BOH, SO, SO₂, PO, PO₂ and SeO, N atom can be quaternized, one or more than one hydrogen atom on the ring is independently optionally substituted with one or more than one substituent described herein.

Heteroaryl can be attached to the main structure via any heteroatom or carbon atom to form a stable compound. Heteroaryl includes, but is not limited to, monocycleformed by 3 to 7 atoms, or bicyclic ring formed by 7 to 10 atoms, or tricyclic ring formed by 10 to 15 atoms. A bicyclic ring containing 7 to 10 atoms can be bicyclic[4,5], [5,5], [5,6] or [6,6] system, tricyclic ring containing 10 to 15 atoms can be tricyclic[5,5,6], [5,7,6] or [6,5,6] system. Depending on the structure, the heteroaryl can be a monovalent or divalent group, i.e. heteroarylene. Examples of heteroaryl include but not limited to 2-furyl, 3-furyl, N-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, 4-methylisoxazol-5-yl, N-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, pyrimidin-5-yl, pyridazinyl(e.g. 3-pyridazinyl), 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, tetrazolyl (e.g. 5-tetrazolyl), triazolyl (e.g. 2-triazolyl and 5-triazolyl), 2-thienyl, 3-thienyl, pyrazolyl (e.g. 2-pyrazolyl), isothiazolyl, 1,2,3-oxadiazolyl, 1,2,5-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,3-triazolyl, 1,2,3-thiodiazolyl, 1,3,4-thiodiazolyl, 1,2,5-thiodiazolyl, 1,3,4-thiadiazole-2-yl, pyrazinyl, pyrazin-2-yl, 1,3,5-triazinyl, benzo[d]thiazol-2-yl, imidazo[1,5-a]pyridin-6-yl, benzimidazolyl, benzoxazolyl, quinoxalinyl, 1,8-naphthyridinyl, benzofuranyl, benzothienyl, benzothiazolyl, indolyl (e.g. 2-indolyl), purinyl, quinolyl (2-quinolinyl, 3-quinolinyl, 4-quinolinyl), isoquinolinyl (e.g, 1-isoquinolyl, 3-isoquinolinyl or 4-isoquinolinyl), tetrahydronaphthyl, benzopyrazolyl, acridinyl, benzimidazolyl, benzoindolyl, benzoisoxazinyl,benzo[4,6]imidazo[1,2-a]pyridinyl,benzo[d]imidazo[2,1-b]thiazolyl, benzofuranyl, benzonaphthofuranyl, benzothiadiazolyl, benzothiazolyl, benzothiophenyl, benzotriazolyl, benzothiopyranyl, benzoxazinyl, benzoxazolyl, benzothiazolyl, β-carbolinyl, carbazolyl, o-naphthyridinyl, dibenzofuranyl, imidazopyridyl, imidazothiazolyl, indazolyl, indolizinyl, indolyl, isobenzothienyl, isoindolinyl, isoquinolinyl, isothiazolidinyl, isothiazolyl, naphthyridinyl, decahydroindolyl, decahydroisoindolyl, oxazolidinedionyl, oxazolidinyl, oxazolopyridinyl, oxazolyl, oxiranyl, tea-diazobenzene, phenanthridinyl, phenanthrolinyl, phenarsazinyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, pyridopyridyl, quinazolinyl, quinoxalinyl, thiophenyl, triazinyl, 2H-pyrrolo[3,4-c]pyridinyl, pyrazolo[2′,1′:2,3]oxazolo[4,5-c]pyridyl, imidazo[2′,1′:2,3]thiazolo[4,5-c]pyridyl, imidazo[2′,1′:2,3]thiazolo[4,5-b]pyridyl, imidazo[2′,1′:2,3]thiazolo[5,4-b]pyridyl, pyrazolo[2′,1′:2,3]thiazolo[4,5-b]pyrazinyl, 1H-benzo[4,5]thieno[2,3-d]imidazolyl, 1-methyl-1H-benzo[4,5]thieno[2,3-d]imidazolyl, imidazo[2′,1′:2,3]thiazolo[4,5-b]pyrazinyl, imidazo[2′,1′:2,3]thiazolo[5,4-b]pyridyl, imidazo[2′,1′:2,3]thiazolo[4,5-c]pyridyl, 1H-benzo[f]imidazo[4,5-b][1,4]thiazepine etc.

The term “aryl” refers to monocyclic, bicyclic, and tricyclic carbon ring system, wherein at least one ring system is aromatic, each ring system contains 3 to 7 atoms, one or more than one hydrogen atom on the ring is independently optionally substituted with one or more than one substituent described herein. The term “aryl” can be used interchangeably with the term “aromatic ring”, e.g. but not limited to, phenyl, naphthyl and anthracene. Depending on the structure, aryl can be a monovalent group or a divalent group, i.e. arylene.

The term “pharmaceutically acceptable salt” refers to a salt formed by a suitable non-toxic organic acid, inorganic acid, organic base or inorganic base with compound I, which retains the biological activity of compound I. The organic acid can be various conventional organic acids in the art and capable of salt formation, preferably selected from the group consisting of methanesulfonic acid, p-toluenesulfonic acid, maleic acid, fumaric acid, citric acid, tartaric acid, malic acid, lactic acid, formic acid, acetic acid, propionic acid, trifluoroacetic acid, oxalic acid, succinic acid, benzoic acid, isethionic acid, naphthalenesulfonic acid and salicylic acid. The inorganic acid can be various conventional inorganic acid in the art and capable of salt formation, preferably selected from the group consisting of hydrochloric acid, sulfuric acid and phosphoric acid. The organic base can be various conventional organic base in the art and capable of salt formation, preferably selected from the group consisting of pyridines, imidazoles, pyrazines, indoles, purines, tertiary amines and anilines. The tertiary amines organic base is preferably triethylamine and/or N,N-diisopropylethylamine. The anilines organic base is preferably N,N-dimethylaniline. The pyridines organic base is preferably selected from the group consisting of pyridine, methylpyridine, 4-dimethylaminopyridine and 2-methyl-5-ethylpyridine. The inorganic base can be various conventional inorganic base in the art and capable of salt formation, preferably selected from the group consisting of alkali metal hydride, alkali metal hydroxide, alkali metal alkoxide, potassium carbonate, sodium carbonate, lithium carbonate, cesium carbonate, potassium bicarbonate and sodium bicarbonate. The alkali metal hydride is preferably sodium hydride and/or potassium hydride. The alkali metal hydroxide is preferably selected from the group consisting of sodium hydroxide, potassium hydroxide and lithium hydroxide. The alkali metal alkoxide is preferably selected from the group consisting of sodium methoxide, sodium ethoxide, potassium tert-butoxide and sodium tert-butoxide.

In the present invention, the “solvate” refers to a substance formed by the compound I and a suitable solvent. The solvent is preferably water or an organic solvent.

Without violating the common sense in the art, the above preferred conditions can be arbitrarily combined, then preferred embodiments of the present invention are obtained.

The reagents and raw materials used in the present invention are commercially available.

The positive and progressive effect of the present invention is that the compound of the present invention exhibits high selectivity and high inhibitory activity with respect to CDK4 and CDK6 at a molecular level, an excellent inhibitory activity with respect to breast cancer cells at a cellular level, and significant inhibition of tumor cell proliferation associated with cyclin-dependent kinase activity at an animal level. The compound also exhibits good stability with respect to liver microsome of human or mouse etc. without significant inhibition of metabolic enzymes, good in vivo absorption in mice and rats, high bioavailability and good druggability.

DETAILED DESCRIPTION

The structure of all the compound of the present invention can be identified by nuclear magnetic resonance (H NMR) and/or mass spectrometry (MS). ¹H NMR chemical shift (δ) is recorded in PPM (10⁻⁶). NMR was determined on a Bruker AVANCE-400 spectrometer.

LC-MS was determined on Agilent 1200 HPLC/6120 mass spectrometer.

The thin-layer silica gel was Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate. Column chromatography generally uses Yantai Huanghai 200-300 mesh silica gel as carrier.

Preparation Embodiment 1

Step 1:

6-Bromo-4-fluoro-1-isopropyl-2-methyl-1H-benzimidazole (200 mg, 0.73765 mmol) (compound represented by formula 1-a), bis(pinacolato)diboron (280 mg, 1.1 mmol), tricyclohexylphosphine (37 mg, 0.1320 mmol), potassium acetate (218 mg, 2.221 mmol) and palladium acetate (19 mg, 0.1148 mmol) were added to dimethyl sulfoxide (2 mL), and the mixture was stirred under nitrogen atmosphere at 90° C. for 2 hours. After cooling to room temperature, the reaction solution was diluted with 10 mL ethyl acetate and filtered. The filtrate was washed with saturated brine, dried over anhydrous sodium sulfate. The organic layer was concentrated and purified by silica gel column chromatography (ethyl acetate/n-hexane 0 to 50%) to give compound 4-fluoro-1-isopropyl-2-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-benzimidazole represented by formula 1-b (180 mg, 0.5657 mmol). LC-MS: m/z: (M+H)⁺=319.2.

Step 2:

2,4-Dichloro-5-fluoropyrimidine (110 mg, 0.65880 mmol) (represented by formula 1-b), 4-fluoro-1-isopropyl-2-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-benzimidazole (180 mg, 0.5657 mmol), (bis(triphenylphosphine))palladium dichloride (30 mg) were added to 2M sodium carbonate solution (1 mL) and ethylene glycol dimethyl ether (3 mL), and the mixture was stirred under nitrogen atmosphere at 85° C. for 2 hours. After cooling to room temperature, the reaction solution was diluted with 10 mL ethyl acetate, washed with saturated brine, and dried over anhydrous sodium sulfate. The organic phase was concentrated and recrystallized from acetonitrile, filtered to give compound 6-(2-chloro-5-fluoro-pyrimidin-4-yl)-4-fluoro-1-isopropyl-2-methyl-benzimidazole represented by formula 1-c (135 mg, 0.4183 mmol). LC-MS: m/z: (M+H)⁺=323.2.

Step 3:

6-(2-Chloro-5-fluoro-pyrimidin-4-yl)-4-fluoro-1-isopropyl-2-methyl-benzimidazole (110 mg, 0.3409 mmol) (represented by formula 1-c), tert-butyl 2-amino-7,8-dihydro-1,6-naphthyridine-6-carboxylate (85 mg, 0.3410 mmol), cesium carbonate (222 mg, 1.1507 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (17 mg, 0.02938 mmol), tris(dibenzylideneacetone)dipalladium (13 mg) were added to 1,4-dioxane (3 mL), and the mixture was stirred under nitrogen atmosphere at 110° C. for 12 hours. After cooling to room temperature, the reaction solution was diluted with 10 mL ethyl acetate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography (dichloromethane/methanol 0-10%) to give tert-butyl 2-((5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)amino)-7,8-dihydro-5H-1,6-naphthyridine-6-carboxylate represented by formula 1-d (110 mg, 0.2054 mmol). LC-MS: m/z: (M+H)⁺=536.2.

Preparation Embodiment 2

Step 1

Methyl 2-methyl nicotinate (1.9 g, 13.0 mmol) (represented by formula 2-a) and trichloroisocyanuric acid (3.7 g, 16.0 mmol) (represented by formula 2-b) were dissolved in dichloromethane (50 mL), and the mixture was stirred at room temperature for 18 hours. Then saturated aqueous sodium bicarbonate solution was added to the reaction solution, the mixture was extracted with dichloromethane (30 mL×2), the dichloromethane layers were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was purified by column chromatography (silica gel, petroleum ether/ethyl acetate=100% to 75%) to give methyl 2-(chloromethyl)nicotinate represented by formula 2-c (1.9 g, 10.3 mmol) as a pale yellow solid. LC-MS: m/z: (M+H)⁺=186.

Step 2:

Methyl 2-(chloromethyl)nicotinate (1.9 g, 10 mmol) (represented by formula 2-c) and m-chloroperoxybenzoic acid (2 g, 11.6 mmol) were dissolved in dichloromethane (50 mL), and the mixture was stirred at room temperature for 16 hours.

Then saturated aqueous sodium bicarbonate solution was added to the reaction solution, the mixture was extracted with dichloromethane (60 mL×2), the dichloromethane layers were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was purified by column chromatography (silica gel, petroleum ether/ethyl acetate=100% to 0%) to give methyl 2-(chloromethyl)nicotinate 1-oxide represented by formula 2-d (15 g, 7.4 mmol) as a pale yellow solid. LC-MS: m/z: (M+H)⁺=202.2.

Step 3:

Methyl 2-(chloromethyl)nicotinate 1-oxide (represented by formula 2-d) (1.9 g, 9.4 mmol) was dissolved in phosphorous oxychloride (9 mL), the mixture was stirred under reflux for 5 hours, then cooled and poured into ice water. Then saturated aqueous sodium bicarbonate solution was added, the mixture was extracted with dichloromethane (60 mL×2), and methylene chloride layers were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was purified by column chromatography (silica gel, petroleum ether/ethyl acetate=100%-0%) to give methyl 6-chloro-2-(chloromethyl)nicotinate represented by formula 2-e (0.88 g, 4.0 mmol) as a pale yellow solid. LC-MS m/z: (M+H)⁺=240.2.

Step 4:

Methyl 6-chloro-2-(chloromethyl) nicotinate (represented by formula 2-e)(70 mg, 0.32 mmol) and N′,N′-dimethylethylenediamine (represented by formula 2-f) (50 mg, 0.57 mmol) were dissolved in tetrahydrofuran (10 mL), and the mixture was stirred at room temperature for 16 hours. The reaction solution was concentrated and purified by thin-layer chromatography (silica gel, dichloromethane/methanol=10/1) to give 2-chloro-6-(2-(dimethylamino)ethyl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one represented by formula 2-g (40 mg, 0.17 mmol) as a pale yellow solid. LC-MS: m/z: (M+H)+=240.2.

Preparation Embodiment 3

Step 1:

6-Bromo-4-fluoro-1-cyclopentyl-2-methyl-benzimidazole (200 mg, 0.6729 mmol) (represented by formula 3-a), bis(pinacolato)diboron (260 mg, 1.1 mmol), tricyclohexylphosphine (37 mg, 0.1320 mmol), potassium acetate (218 mg, 2.221 mmol) and palladium acetate (19 mg, 0.1148 mmol) were added to dimethyl sulfoxide (2 mL), and the mixture was stirred under nitrogen atmosphere at 90° C. for 2 hours. After cooling to room temperature, the reaction solution was diluted with 10 mL ethyl acetate and filtered. The filtrate was washed with saturated brine, dried over anhydrous sodium sulfate. The organic layer was concentrated and purified by silica gel column chromatography (ethyl acetate/n-hexane 0-50%) to give 4-fluoro-1-cyclopentyl-2-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-benzimidazole represented by formula 3-b (180 mg, 0.523 mmol). LC-MS: m/z: (M+H)⁺=345.2.

Step 2:

2,4-Dichloro-5-fluoropyrimidine (110 mg, 0.65880 mmol), 4-fluoro-1-cyclopentyl-2-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-benzimidazole (180 mg, 0.523 mmol) (represented by formula 3-b), (bis(triphenylphosphine)palladium dichloride (30 mg) were added to 2M sodium carbonate solution (1 mL) and ethylene glycol dimethyl ether (3 mL), and the mixture was stirred under nitrogen atmosphere at 85° C. for 2 hours. After cooling to room temperature, the reaction solution was diluted with 10 mL ethyl acetate, washed with saturated brine, and dried over anhydrous sodium sulfate. The organic phase was concentrated and recrystallized from acetonitrile to give 6-(2-chloro-5-fluoro-pyrimidine-4-yl)-4-fluoro-1-cyclopentyl-2-methyl-benzimidazole represented by formula 3-c (135 mg, 0.37 mmol). LC-MS: m/z: (M+H)⁺=349.8

Step 3:

5-(2-Chloro-5-fluoropyrimidin-4-yl)-1-cyclopentyl-7-fluoro-2-methyl-1H-benzo[d]imidazole) (360 mg, 1.03 mmol) (represented by formula 3-c) was dissolved in 5 mL dioxane, tert-butyl 2-amino-7,8-dihydro-1,6-naphthyridine-6(5H)-methylcarboxylate (257 mg, 1.03 mmol), tris(dibenzylideneacetone)dipalladium (94 mg, 0.10 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (119 mg, 0.21 mmol) and cesium carbonate (504 mg, 1.55 mmol) were added, and the mixture was stirred under argon atmosphere at 110° C. for 18 hours. The reaction solution was filtered and evaporated under reduced pressure. The residue was purified by silica gel column chromatography to give the compound represented by formula 3-d as a yellow solid 340 mg, yield 82%. LC-MS: m/z: (M+H)⁺=548.2.

Preparation Embodiment 4

Step 1:

Cyclopropylamine (30 g, 525.5 mmol) and diisopropylacetamide (80 g, 790.6 mmol) were dissolved in dichloromethane (200 mL), the mixture was cooled to 0° C. and acetic anhydride was slowly added dropwise (115 g, 1126 mmol). After the completion of the addition, the mixture was stirred at room temperature for 16 hours, evaporated under reduced pressure, then ethyl acetate and potassium carbonate (144 g, 1050 mmol) were added. The mixture was stirred at room temperature for 16 hours, filtered, and evaporated under reduced pressure to give 55 g N-cyclopropylacetamide represented by formula 4-c.

Step 2:

N-cyclopropylacetamide (54 g, 544.7 mmol), 4-bromo-2,6-difluoroaniline (54.6 g, 263 mmol) and N,N-diisopropylethylamine (50.8 g, 393 mmol) were dissolved in toluene (250 mL), then phosphorus oxychloride (40.3 g, 263 mmol) was slowly added. The mixture was stirred at 100° C. for 16 hours, evaporated under reduced pressure, cooled to room temperature, then dichloromethane was added, washed with saturated sodium bicarbonate solution, and the organic layer was dried over anhydrous sodium sulfate, concentrated, slurried with ethyl acetate, filtered, and the solid was dried to give 43.6 g N′-(4-bromo-2,6-difluoro-phenyl)-N-cyclopropyl-acetamidine represented by formula 4-d. LC-MS m/z: (M+H)⁺=289.1, 291.1.

Step 3:

N′-(4-bromo-2,6-difluoro-phenyl)-N-cyclopropyl-acetamidine (43.6 g, 151 mmol) was dissolved in N,N-dimethylformamide (300 mL), potassium tert-butoxide (28 g, 249.5 mmol) was added slowly, and the mixture was stirred at 100° C. for 3 hours, cooled to room temperature. Then 1.5 L water was added, the mixture was filtered, the solid was washed with water and distilled under reduced pressure to remove water, then slurried with a mixed solvent of dichloromethane/petroleum ether=1/2, and filtered to give 35 g 6-bromo-1-cyclopropyl-4-fluoro-2-methyl-benzimidazole represented by formula 4-f. LC-MS m/z: (M+H)⁺=269.1, 271.1.

Step 4:

6-Bromo-1-cyclopropyl-4-fluoro-2-methyl-benzimidazole (15 g, 55.7 mmol), bis(pinacolato)diboron (16 g, 63 mmol), potassium acetate (10 g, 102 mmol), tricyclohexylphosphine (1.5 g, 5.3 mmol) and palladium acetate (1 g, 4.45 mmol) were dissolved in DMSO (60 mL), and the mixture was stirred under argon atmosphere at 80° C. for 16 hours. The reaction solution was cooled to room temperature, filtered, diluted with water and extracted with dichloromethane. The organic layer was dried over anhydrous sodium sulfate and concentrated to give a crude product, which was slurried with ethyl acetate. Partial product was collected by filtration, and the filtrate was concentrated and purified by column chromatography (petroleum ether/ethyl diacid=100% to 25%) to obtain partial product, the product was combined to give 5 g 1-cyclopropyl-4-fluoro-2-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-benzimidazole represented by formula 4-e. LC-MS m/z: (M+H)⁺=317.2.

Step 5:

1-Cyclopropyl-4-fluoro-2-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-benzimidazole (5 g, 15.8 mmol), 2,4-dichloro-5-fluoro-pyrimidine (3 g, 18.0 mmol), sodium carbonate (3 g, 28.3 mmol) and dichlorobis(triphenylphosphine)palladium (550 mg, 0.83 mmol) were dissolved in a mixed solution of ethylene glycol dimethyl ether (70 mL) and water (10 mL), the mixture was stirred under argon atmosphere at 80° C. for 16 hours. The reaction solution was cooled to room temperature, filtered, and the ethylene glycol dimethyl ether was distilled off under reduced pressure. The residue was diluted with water and extracted with dichloromethane. The organic layer was dried over anhydrous sodium sulfate, and concentrated to give a crude product, which was slurried with acetonitrile, filtered and dried to give 4.5 g 6-(2-chloro-5-fluoro-pyrimidin-4-yl)-1-cyclopropyl-4-fluoro-2-methyl-benzimidazole represented by 4-g. LC-MS m/z: (M+H)⁺=321.2.

Step 6:

6-(2-Chloro-5-fluoro-pyrimidin-4-yl)-1-cyclopropyl-4-fluoro-2-methyl-benzimidazole (4.5 g, 14 mmol), tert-butyl 2-amino-7,8-dihydro-5H-1,6-naphthyridine-6-carboxylate (3.8 g, 15 mmol), cesium carbonate (9 g, 27.6 mmol), tris(dibenzylideneacetone)dipalladium (900 mg, 0.98 mmol) and 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (500 mg, 0.86 mmol) were dissolved in dioxane (100 mL), the mixture was stirred under argon atmosphere at 90° C. for 16 hours, then distilled under reduced pressure to remove the solvent, diluted with a mixed solvent of dichloromethane and methanol, filtered. The filtrate was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (dichloromethane/methanol=100% to 90%) to give 7 g tert-butyl 2-((5-fluoro-4-(7-fluoro-3-cyclopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)amino)-7,8-dihydro-5H-1,6-naphthyridine-6-carboxylate represented by 4-h. LC-MS m/z: (M+H)⁺=534.2.

Preparation Embodiment 5

The mixture of tert-butyl 2-amino-7-methyl-7,8-dihydro-5H-1,6-naphthyridine-6-carboxylate and tert-butyl 2-amino-5-methyl-7,8-dihydro-5H-1,6-naphthyridine-6-carboxylate (represented by formula 5-a and 5-b, supplied by KERMANDA) (3.0 g, 11.4 mmol) was isolated via chiral liquid phase preparation to give four isomers, which were tert-butyl 2-amino-5-methyl-7,8-dihydro-5H-1,6-naphthyridine-6-carboxylate (Peak 1, 0.5658 g), tert-butyl 2-amino-5-methyl-7,8-dihydro-5H-1,6-naphthyridine-6-carboxylate (Peak 3, 0.6884 g), tert-butyl 2-amino-7-methyl-7,8-dihydro-5H-1,6-naphthyridine-6-carboxylate (Peak 2, 0.4281 g), tert-butyl 2-amino-7-methyl-7,8-dihydro-5H-1,6-naphthyridine-6-carboxylate (Peak 4, 0.6460 g). LC-MS m/z: (M+H)⁺=264.2.

Preparation Embodiment 6

Step 1:

DIPEA (1.757 g, 13.59 mmol) and 2-bromoethanol (1.446 g, 11.57 mmol) were added to a suspension of N-(4-(3-tert-butyl-7-fluoro-2-methyl-benzimidazol-5-yl)-5-fluoro-pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine (1.3 g, 2.892 mmol) (represented by formula I-68) in anhydrous DMF (22 mL) under argon atmosphere at room temperature. The reaction mixture was stirred at 85° C. for 12 hours. TLC monitored that most of the raw materials had already reacted. The reaction mixture was cooled to room temperature and diluted with 20 mL water. The precipitated solid was filtered and the filter cake was washed with water (10 mL×3) and dried in vacuo to give a yellow solid 2-(2-((4-(3-tert-butyl-7-fluoro-2-methyl-benzoimidazol-5-yl)-5-fluoro-pyrimidin-2-yl)amino)-7,8-dihydro-5H-1,6-naphthyridin-6-yl)ethanol represented by formula 3-b (1.1 g, 2.229 mmol) as a pale yellow solid.

¹H NMR (400 MHz, Chloroform-d) δ 8.40 (d, J=3.8 Hz, 1H), 8.39 (d, J=1.3 Hz, 1H), 8.23 (d, J=8.4 Hz, 1H), 7.93 (s, 1H), 7.75 (d, J=11.2 Hz, 1H), 7.36 (d, J=8.5 Hz, 1H), 3.78-3.72 (m, 2H), 3.70 (s, 2H), 2.95 (dd, J=10.7, 4.6 Hz, 4H), 2.88 (s, 3H), 2.80-2.75 (m, 2H), 1.91 (s, 9H).

LC-MS m/z:(M+H)⁺=494.2.

Step 2:

Thionyl chloride (7.954 g, 66.86 mmol) was added to a suspension of 2-(2-((4-(3-tert-butyl-7-fluoro-2-methyl-benzimidazol-5-yl)-5-fluoro-pyrimidin-2-yl)amino)-7,8-dihydro-5H-1,6-naphthyridin-6-yl)ethanol (1.1 g, 2.229 mmol) (represented by formula 6-a) in dichloromethane (40 mL) at room temperature under argon atmosphere. The mixture was stirred at room temperature for 16 hours. LCMS monitored the reaction was complete. The solvent was removed by distillation under reduced pressure. The solid residue was dissolved in 100 mL mixed solvent of dichloromethane and methanol (10:1, v/v). The organic phase was washed to neutral with saturated aqueous sodium bicarbonate solution, then washed with saturated brine (50 mL×2). The separated organic phase was evaporated to dryness on a rotary evaporator, the residue was purified by biotage preparative liquid chromatography (methanol/dichloromethane=1% to 5%, v/v, 10 g silica gel column) to give yellow solid N-(4-(3-tert-butyl-7-fluoro-2-methyl-benzimidazol-5-yl)-5-fluoro-pyrimidin-2-yl)-6-(2-chloroethyl)-7,8-dihydro-5H-1,6-naphthyridin-2-amine (600 mg, 1.172 mmol) as a pale yellow solid.

¹H NMR (400 MHz, Chloroform-d) δ 8.41-8.36 (m, 2H), 8.22 (d, J=8.5 Hz, 1H), 7.91 (s, 1H), 7.75 (d, J=11.4 Hz, 1H), 7.35 (d, J=8.4 Hz, 1H), 3.72-3.67 (m, 4H), 3.00-2.90 (m, 6H), 2.87 (s, 3H), 1.91 (s, 9H).

LC-MS m/z:(M+H)⁺=512.2.

Embodiment 3

N-(5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridine-2-amine (70 mg, 0.1607 mmol)(represented by formula I-1), 2-bromopropane (40 mg, 0.32523 mmol), N,N-diisopropyl ethylamine (50 mg) were added to 1 mL DMF, and the mixture was stirred at 90° C. for 12 hours. The reaction solution was concentrated and purified by silica gel column chromatography (dichloromethane/methanol 0 to 10%) to give N-(5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)-6-isopropyl-7,8-dihydro-5H-1,6-naphthyridin-2-amine represented by I-3 (30 mg, 0.06281 mmol). ¹H-NMR (400 MHz, DMSO-d₆) δ 9.90 (s, 1H), 8.68 (d, 1H, J=3.6 Hz), 8.32 (d, 1H, J=1.2 Hz), 8.06 (d, 1H, J=8.4 Hz), 7.69 (d, 1H, J=12.4 Hz), 7.48 (d, 1H, J=1.2 Hz), 4.82-4.89 (m, 1H), 3.63 (s, 2H), 2.81 (br, 4H), 1.97-2.04 (m, 1H), 1.64 (d, 6H, J=6.8 Hz), 1.08 (d, 6H, J=6.0 Hz). LC-MS: m/z: (M+H)⁺=478.2.

Embodiment 4

Step 1:

N-(5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridine-2-amine (80 mg, 0.1837 mmol) (represented by formula I-1), N-boc-sarcosine (46 mg, 0.24312 mmol) (represented by formula I-3-a), HOBt (25 mg, 0.18502 mmol), EDCI (36 mg, 0.18779 mmol) were added to 1 mL DMF, and the mixture was stirred at room temperature for 12 hours. The reaction solution was concentrated and purified by silica gel column chromatography (dichloromethane/methanol 0 to 10%) to give tert-butyl N-(2-(2-((5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)amino)-7,8-dihydro-5H-1,6-naphthyridine-6-yl)-2-oxoethyl)-N-methyl-carbamate represented by formula I-4-b (50 mg, 0.08241 mmol). LC-MS: m/z: (M+H)⁺=607.2.

Step 2:

Tert-butyl N-(2-(2-((5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)amino)-7,8-dihydro-5H-1,6-naphthyridin-6-yl)-2-oxoethyl)-N-methylcarbamate (50 mg, 0.08241 mmol) (represented by formula I-4-b) was added to 2M HCl/MeOH solution (2 mL), and the mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated to give 1-(2-((5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidine-2-yl)amino)-7,8-dihydro-5H-1,6-naphthyridin-6-yl)-2-(methylamino)ethanone represented by formula I-4 (30 mg, 0.05922 mmol). ¹H-NMR (400 MHz, CD₃OD) δ 8.96-8.97 (m, 1H), 8.62 (s, 1H), 8.22-8.26 (m, 2H), 7.56 (dd, 1H, J=8.8, 2.8 Hz), 5.16-5.23 (m, 1H), 4.87 (s, 1H), 4.78 (s, 1H), 4.32 (s, 1H), 4.27 (s, 1H), 4.08 (t, 1H, J=6.0 Hz), 3.92 (t, 1H, J=6.0 Hz), 3.37-3.38 (m, 2H), 3.24-3.27 (m, 2H), 3.02 (s, 3H), 2.82 (d, 3H, J=2.8 Hz), 1.84 (d, 6H, J=6.8 Hz). LC-MS: m/z: (M+H)⁺=507.2.

Embodiment 5

N-(5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridine-2-amine (80 mg, 0.1837 mmol)(represented by formula I-1), N,N-dimethylglycine (20.5 mg, 0.199 mmol), HOBt (25 mg, 0.18502 mmol), EDCI (36 mg, 0.18779 mmol) were added to 1 mL DMF, and the mixture was stirred at room temperature for 12 hours. The reaction solution was concentrated and purified by silica gel column chromatography (dichloromethane/methanol 0 to 10%) to give 2-(dimethylamino)-1-(2-((5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)amino)-7,8-dihydro-5H-1,6-naphthyridine-6-yl)ethanone represented by formula I-5 (10 mg, 0.01921 mmol). ¹H-NMR (400 MHz, CD₃OD) δ 8.50 (d, 1H, J=3.6 Hz), 8.26-8.30 (m, 2H), 7.78 (d, 1H, J=12 Hz), 7.56 (d, 1H, J=8.4 Hz), 4.86-4.90 (m, 2H), 4.86 (s, 2H), 4.71 (s, 1H), 3.92 (t, 2H, J=5.6 Hz), 3.00 (t, 1H, J=5.6 Hz), 2.92 (t, 1H, J=5.6 Hz), 2.69 (s, 3H), 2.35 (s, 3H), 2.34 (s, 3H), 1.74 (d, 6H, J=7.2 Hz). LC-MS: m/z: (M+H)⁺=521.2.

Embodiment 6

N-(5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridine-2-amine (80 mg, 0.1837 mmol)(represented by formula I-1), 2-methoxyacetic acid (18 mg, 0.1998 mmol), HOBt (25 mg, 0.18502 mmol), EDCI (36 mg, 0.18779 mmol) were added to 1 mL DMF and the mixture was stirred at room temperature for 12 hours. The reaction solution was concentrated and purified by silica gel column chromatography (dichloromethane/methanol 0 to 10%) to give 1-(2-((5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)amino)-7,8-dihydro-5H-1,6-naphthyridin-6-yl)-2-methoxyethanone (60 mg, 0.1182 mmol). ¹H-NMR (400 MHz, CD₃OD) δ 8.54 (d, 1H, J=3.6 Hz), 8.33 (s, 1H), 8.23-8.27 (m, 1H), 7.76 (d, 1H, J=11.6 Hz), 7.57 (d, 1H, J=8.4 Hz), 4.87-4.96 (m, 1H), 4.70 (s, 1H), 4.67 (s, 1H), 4.30 (s, 1H), 4.28 (s, 1H), 3.92 (t, 1H, J=5.6 Hz), 3.82 (t, 1H, J=5.6 Hz), 3.46 (s, 1.8H), 3.44 (s, 1.2H), 2.98 (t, 1H, J=5.6 Hz), 2.93 (t, 1H, J=5.6 Hz), 2.71 (s, 3H), 1.73 (d, 6H, J=7.6 Hz). LC-MS: m/z: (M+H)⁺=508.2.

Embodiment 8

N-(5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridine-2-amine (80 mg, 0.1837 mmol)(represented by formula I-1), aqueous formaldehyde (20 mg), sodium triacetoxyborohydride (128 mg) were added to 3 mL dioxane, the mixture was stirred at room temperature for 12 hours. The reaction solution was concentrated and purified by silica gel column chromatography (dichloromethane/methanol 0 to 10%) to give N-(5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzoimidazol-5-yl)pyrimidin-2-yl)-6-methyl-7,8-dihydro-5H-1,6-naphthyridin-2-amine represented by formula I-8 (40 mg, 0.08899 mmol). ¹H-NMR (400 MHz, CD₃OD) δ 8.54 (d, 1H, J=4 Hz), 8.33 (d, 1H, J=1.2 Hz), 8.22 (d, 1H, J=8.4 Hz), 7.52 (d, 1H, J=8.4 Hz), 4.85-4.96 (m, 1H), 3.75 (s, 2H), 2.98-3.03 (m, 4H), 2.71 (s, 3H), 2.59 (s, 3H), 1.33 (d, 6H, J=7.2 Hz). LC-MS: m/z: (M+H)⁺=450.2.

Embodiment 9

N-(5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridine-2-amine (80 mg, 0.1837 mmol)(represented by formula I-1), 3-hydroxypropionic acid (18 mg, 0.1998 mmol), HOBt (25 mg, 0.18502 mmol), EDCI (36 mg, 0.18779 mmol) were added to 1 mL DMF and the mixture was stirred at room temperature for 12 hours. The reaction solution was concentrated and purified by silica gel column chromatography (dichloromethane/methanol 0 to 10%) to give 1-(2-((5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)amino)-7,8-dihydro-5H-1,6-naphthyridin-6-yl)-3-hydroxyacetone (8 mg, 0.01576 mmol). ¹H-NMR (400 MHz, DMSO-d₆) δ 10.05 (d, 1H, J=10 Hz), 8.70 (d, 1H, J=4 Hz), 8.32 (s, 1H), 8.13 (m, 1H), 7.71 (d, 1H, J=12.4 Hz), 7.60-7.63 (m, 1H), 4.83-4.89 (m, 1H), 4.69 (s, 1H)), 4.62 (s, 1H), 3.81-3.83 (m, 2H), 3.68-3.69 (m, 2H), 2.91 (t, 1H, J=5.6 Hz), 2.80 (t, 1H, J=5.6 Hz), 2.66 (s, 3H), 2.60-2.63 (m, 2H). LC-MS: m/z:(M+H)⁺=508.2.

Embodiment 10

Step 1:

N-(5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl]-5,6,7,8-tetrahydro-1,6-naphthyridine-2-amine (80 mg, 0.1837 mmol)(represented by formula I-1), pyridine (120 mg, 1.517 mmol), methyl oxalyl chloride (50 mg, 0.40813 mmol) (represented by formula I-10-a) were added to 10 mL THE and the mixture was stirred at room temperature for 12 hours. The reaction solution was concentrated and purified by silica gel column chromatography (dichloromethane/methanol 0 to 10%) to give methyl 2-(2-((5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)-(2-methoxy-2-oxoacetyl)amino)-7,8-dihydro-5H-1,6-naphthyridin-6-yl)-2-oxoacetate represented by formula I-10-b (100 mg, 0.1646 mmol). LC-MS: m/z: (M+H)+=608.2.

Step 2:

Methyl 2-(2-((5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)-(2-methoxy-2-oxoacetyl)amino)-7,8-dihydro-5H-1,6-naphthyridin-6-yl)-2-oxoacetate (100 mg, 0.1646 mmol) (represented by formula I-10-b), lithium hydroxide (40 mg) and water (0.5 mL) were added to 3 mL methanol and the mixture was stirred at room temperature for 12 hours. The reaction solution was concentrated and purified by silica gel column chromatography (dichloromethane/methanol 0 to 10%) to give methyl 2-(2-((5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl]amino]-7,8-dihydro-5H-1,6-naphthyridin-6-yl]-2-oxoacetate represented by formula I-10 (70 mg, 0.1342 mmol). ¹H-NMR (400 MHz, DMSO-d₆) δ 10.10 (d, 1H, J=7.2 Hz), 8.69 (d, 1H, J=4 Hz), 8.32 (d, 1H, J=1.2 Hz), 8.14-8.18 (m, 1H), 7.61-7.72 (m, 2H), 4.82-4.89 (m, 1H), 4.68 (s, 1H), 4.60 (s, 1H), 3.87-3.88 (m, 4H), 3.75 (t, 1H, J=5.6 Hz), 2.88-2.92 (m, 2H), 2.65 (s, 3H), 1.64 (d, 6H, J=6.8 Hz). LC-MS: m/z:(M+H)⁺=522.2.

Embodiment 12

N-(5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine (80 mg, 0.18 mmol) (represented by formula I-1), 2,2,2-trifluoroethyl methanesulfonate (60 mg, 0.26 mmol) and diisopropylethylamine (60 mg, 0.46 mmol) were dissolved in N,N-dimethylformamide (3 mL) and the mixture was stirred at 90° C. for 16 hours. Then the reaction solution was cooled to room temperature and filtered, the residue was dissolved in dichloromethane, treated with ultrasound for 30 minutes, filtered, rinsed with dichloromethane, and the residue was dissolved in methanol, treated with ultrasound for 30 minutes, then filtered to give N-(5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)-6-(2,2,2-trifluoroethyl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine represented by formula I-12 (45 mg) as a pale yellow solid. ¹H-NMR (400 MHz, DMSO-d₆) δ 9.97 (s, 1H), 8.68 (d, 1H, J=4.0 Hz), 8.31 (d, 1H, J=1.2 Hz), 8.08 (d, 1H, J=8.4 Hz), 7.71 (d, 1H, J=12.4 Hz), 7.48 (d, 1H, J=8.4 Hz), 4.83-4.88 (m, 1H), 3.82 (s, 2H), 3.40 (q, 2H, J=10.4 Hz), 3.04 (t, 2H, J=6.0 Hz), 2.86 (t, 2H, J=6.0 Hz), 2.66 (s, 3H), 1.64 (d, 6H, J=6.8 Hz). LC-MS: m/z: (M+H)⁺=518.2

Embodiment 13

Step 1:

6-(2-Chloro-5-fluoro-pyrimidin-4-yl)-4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazole (100 mg, 0.31 mmol) (represented by formula 1-c), tert-butyl 2-amino-7,8-dihydropyrido[4,3-d]pyrimidin-6(5H)-carboxylate (100 mg, 0.4 mmol) (represented by formula I-13-a), tris(dibenzylideneacetone)dipalladium (50 mg, 0.05 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (30 mg, 0.05 mmol) and cesium carbonate (326 mg, 1 mmol) were dissolved in 1,4-dioxane (6 mL) and the mixture was stirred under argon atmosphere at 100° C. for 18 hours. Then the reaction solution was concentrated and purified by column chromatography (dichloromethane/methanol: 0% to 10%) to give tert-butyl 2-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)-7,8-dihydropyrido[4,3-d]pyrimidine-6(5H)-carboxylate (45 mg) as pale yellow solid. LC-MS: m/z: (M+H)⁺=537.2.

Step 2:

Tert-butyl 2-((5-Fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)-7,8-dihydropyrido[4,3-d]pyrimidine-6(5H)-carboxylate (45 mg, 0.08 mmol) (represented by formula I-13-b) was dissolved in tetrahydrofuran (2 mL), hydrochloric acid (4M) in tetrahydrofuran was added, the mixture was stirred at room temperature for 1 hour, then concentrated to give N-(5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-hydrobenzo[d]imidazol-6-yl)pyrimidin-2-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-amine hydrochloride (40 mg) represented by 1-13 as a pale yellow solid. ¹H-NMR (400 MHz, DMSO-d₆) δ 10.67 (s, 1H), 9.59 (s, 1H), 8.78 (d, 1H, J=3.6 Hz), 8.53 (s, 1H), 8.39 (s, 1H), 7.92 (d, 1H, J=11.6 Hz), 4.90-4.94 (m, 1H), 4.29 (t, 2H, J=4.8 Hz), 3.51 (q, 2H, J=6.0 Hz), 3.06 (t, 2H, J=6.4 Hz), 2.75 (s, 3H), 1.65 (d, 6H, J=6.8 Hz). LC-MS m/z:(M+H)⁺=437.2.

Embodiment 14

N-(5-Fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-amine hydrochloride (30 mg, 0.06 mmol) (represented by formula I-13), N,N-dimethylglycine (8 mg, 0.08 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (30 mg, 0.16 mmol), 1-hydroxybenzotriazole (30 mg, 0.22 mmol) and diisopropylacetamide (100 mg, 0.78 mmol) were dissolved in 1,4-dioxane (4 mL) and the mixture was stirred at room temperature for 16 hours. Then the reaction solution was concentrated, purified with TLC (DCM/CH₃OH=10/1), followed by column chromatography (C18, H₂O/CH₃OH=100% to 5%) to give 2-dimethylamino-1-(2-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)-7,8-dihydropyrido[4,3-d]pyrimidin-6(5H)-yl)acetamide represented by formula I-14 (9 mg) as pale yellow solid. ¹H-NMR (400 MHz, CDCl₃) δ 8.54-8.55 (m, 1H), 8.44 (s, 1H), 8.22-8.25 (m, 1H), 8.15 (s, 1H), 7.90-7.95 (m, 1H), 4.73-4.85 (m, 3H), 3.97 (t, 2H, J=6.0 Hz), 3.24-3.26 (m, 2H), 2.98-3.08 (m, 2H), 2.72 (s, 3H), 2.30-2.33 (m, 6H), 1.73 (d, 6H, J=6.8 Hz). LC-MS: m/z: (M+H)⁺=522.3.

Embodiment 15

Step 1:

6-(2-Chloro-5-fluoro-pyrimidin-4-yl)-4-fluoro-1-isopropyl-2-methyl-benzimidazole (500 mg, 1.55 mmol) (represented by formula 1-c), aqueous ammonia (5 mL, 35%) and 1,4-dioxane (5 mL) were added to a sealed tube and the mixture was stirred at 100° C. for 16 hours, then concentrated and diluted with dichloromethane (30 mL). After 30 minutes under ultrasound, the mixture was filtered, and the residue was rinsed with dichloromethane and dried to give 5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-amine represented by formula I-15-a (280 mg, 0.92 mmol) as a pale yellow solid. LC-MS m/z: (M+H)⁺=304.2.

Step 2:

2-Chloro-6-(2-(dimethylamino)ethyl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one (40 mg, 0.17 mmol) (represented by formula 2-g), 5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidine-2-amine (60 mg, 0.19 mmol) (represented by formula I-15-a), tris(dibenzylideneacetone)dipalladium (30 mg, 0.03 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (20 mg, 0.04 mmol) and cesium carbonate (120 mg, 0.37 mmol) were dissolved in 1,4-dioxane (6 mL) and the mixture was stirred under argon atmosphere at 100° C. for 18 hours, then concentrated and purified by column chromatography (dichloromethane/methanol 0% to 10%) to give 6-(2-(dimethylamino)ethyl)-2-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one (45 mg, 0.09 mmol) represented by formula I-15 as a pale yellow solid. ¹H-NMR (400 MHz, CDCl₃) δ 8.62 (d, 1H, J=8.8 Hz), 8.44 (s, 1H), 8.50 (d, 1H, J=3.6 Hz), 8.40 (s, 1H), 8.24 (d, 1H, J=1.2 Hz), 8.09 (d, 1H, J=11.6 Hz), 7.80 (d, 1H, J=11.6 Hz), 4.74-4.79 (m, 1H), 4.53 (s, 2H), 3.89 (t, 2H, J=7.8 Hz), 2.86 (t, 2H, J=6.4 Hz), 2.72 (s, 3H), 2.51 (s, 6H), 1.75 (d, 6H, J=6.8 Hz). LC-MS m/z: (M+H)⁺=507.9

Embodiment 17

Step 1:

N-(5-fluoro-4-(7-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-5-yl)pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine (80 mg, 0.18 mmol) (represented by formula I-1), N-boc-glycine (39 mg, 0.22 mmol) (represented by formula I-17-a), HOBt (30 mg, 0.22 mmol), EDCI (42 mg, 0.22 mmol) were added to 1 mL DMF and the mixture was stirred at room temperature for 12 hours. The reaction solution was concentrated and purified by silica gel column chromatography (dichloromethane/methanol 0-10%) to give the compound represented by formula I-17-b as a yellow solid 43 mg, yield 39%. ¹H-NMR (400 MHz, CDCl₃) δ 8.45 (d, 1H, J=3.6 Hz), 8.34 (dd, 1H, J=8.5, 3.3 Hz), 8.17-8.21 (m, 2H), 7.79 (d, 1H, J=11.7 Hz), 7.48 (d, 0.66H, J=8.6 Hz), 7.43 (d, 0.44H, J=8.6 Hz), 5.57 (s, 1H), 4.76 (s, 1H), 4.71-4.79 (m, 1H), 4.56 (s, 1H), 4.10 (t, 2H, J=3.2 Hz), 3.98 (t, 1H, J=6.0 Hz), 3.75 (t, 1H, J=5.9 Hz), 3.00 (t, 1H, J=6.0 Hz), 2.96 (t, 1H, J=6.0 Hz), 2.71 (s, 3H), 1.73 (d, J=6.9 Hz, 6H), 1.48 (s, 9H). LC-MS: m/z:(M+H)⁺=593.2.

Step 2:

Tert-butyl (2-(2-((5-Fluoro-4-)₇-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-5-yl)pyridin-2-yl)amino)-7,8-dihydro-1,6-naphthyridine-6(5H)-yl)-2-oxoethyl)carbamate (43 mg, 0.23 mmol) (represented by formula I-17-b) was dissolved in 5 mL dioxane, 4N HCl in dioxane (1 mL) was added dropwise, and the mixture was stirred at room temperature for 4 hours. The solvent was evaporated under reduced pressure, saturated sodium bicarbonate solution was added, and the mixture was extracted with dichloromethane. The solvent was evaporated under reduced pressure to give 21 mg the compound represented by formula I-17 as a yellow solid, yield 59%. ¹H-NMR (400 MHz, CD₃OD) δ 8.94 (s, 1H), 8.58 (s, 1H), 8.20 (t, 2H, J=9.3 Hz), 7.56 (d, 1H, J=8.8 Hz), 5.14-5.15 (m, 1H), 4.78 (s, 2H), 4.17 (s, 1H), 4.14 (s, 1H), 4.08 (s, 1H), 3.92 (s, 1H), 3.24 (s, 2H), 2.97 (s, 3H), 1.83 (d, 6H, J=6.9 Hz). LC-MS: m/z:(M+H)⁺=493.2.

Embodiment 19

N-(5-Fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridine-2-amine (200 mg, 0.46 mmol)(represented by formula I-1), methyl bromoacetate (281 mg, 1.837 mmol), N,N-diisopropylethylamine (178 mg) were added to 10 mL 1,4-dioxane, and the mixture was stirred at 90° C. for 12 hours. The reaction solution was concentrated and purified by silica gel column chromatography (dichloromethane/methanol 0 to 10%) to give methyl 2-(2-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)acetate represented by formula I-19 (130 mg, 0.256 mmol). ¹H-NMR (400 MHz, CD₃OD) δ 8.53 (d, J=3.9 Hz, 1H), 8.32 (s, 1H), 8.16 (d, J=8.5 Hz, 1H), 7.75 (d, J=12.2 Hz, 1H), 7.46 (d, J=8.5 Hz, 1H), 4.89-4.95 (m, 1H), 3.77-3.81 (m, 2H), 3.78 (s, 3H), 3.62 (s, 0.5H), 3.51 (s, 1.5H), 2.97-3.02 (m, 4H), 2.70 (s, 3H), 1.73 (d, 6H, J=6.9 Hz). LC-MS m/z: (M+H)⁺=508.2.

Embodiment 20

Methyl 2-(2-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)acetate (100 mg, 0.197 mmol) (represented by formula I-19), LiOH (42 mg), H₂O (10 mL) were added to MeOH (10 mL), the mixture was stirred at 75° C. for 1 hour. The reaction solution was filtered and washed to give 2-(2-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)acetic acid represented by formula I-20 (70 mg, 0.14 mmol). ¹H-NMR (400 MHz, DMSO-d₆) δ 10.00 (s, 1H), 8.69 (d, J=4.3 Hz, 1H), 8.32 (s, 1H), 8.09 (d, J=8.7 Hz, 1H), 7.71 (d, J=11.7 Hz, 1H), 7.50 (d, J=8.5 Hz, 1H), 4.83-4.88 (m, 1H), 3.87 (s, 2H), 3.04 (s, 2H), 2.90 (s, 2H), 2.66 (s, 3H), 2.00-2.04 (m, 2H), 1.64 (s, 6H, 6.8 Hz). LC-MS: m/z: (M+H)⁺=495.2.

Embodiment 21

Tert-butyl 2-((4-(1-Cyclopentyl-7-fluoro-2-methyl-1H-benzo[d]imidazol-5-yl)-5-fluoropyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridine-6(5H)-carboxylate (340 mg, 0.61 mmol) (represented by formula 3-d) were dissolved in 5 mL dioxane, 4N HCl in dioxane (1 mL) was added dropwise and the mixture was stirred at room temperature for 4 hours. Solvent was evaporated under reduced pressure, saturated sodium bicarbonate solution was added, the mixture was extracted with dichloromethane, and the solvent was evaporated under reduced pressure to give 230 mg the compound represented by formula I-21 as a yellow solid, yield 82%. ¹H-NMR (400 MHz, DMSO-d₆) δ 9.83 (s, 1H), 8.67 (d, 1H, J=3.8 Hz), 8.12 (s, 1H), 8.02 (d, 1H, J=7.6 Hz), 7.73 (d, 1H, J=12.1 Hz), 7.39-7.45 (m, 1H), 5.06-4.90 (m, 1H), 4.41 (s, 1H), 3.82 (s, 1H), 3.58 (s, 1H), 3.18 (s, 2H), 3.03 (s, 1H), 2.66 (s, 3H), 2.11-2.20 (m, 4H), 1.96-2.01 (m, 2H), 1.83-1.69 (m, 2H). LC-MS: m/z: (M+H)⁺=461.9.

Embodiment 22

Step 1:

N-(5-Fluoro-4-(7-fluoro-3-cyclopentyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl]-5,6,7,8-tetrahydro-1,6-naphthyridine-2-amine (90 mg, 0.195 mmol) (represented by formula I-21), N-boc-sarcosine (46 mg, 0.24312 mmol), HOBt (25 mg, 0.18502 mmol), EDCI (36 mg, 0.18779 mmol) were added to 1 mL DMF and the mixture was stirred at room temperature for 12 hours. The reaction solution was concentrated and purified by silica gel column chromatography (dichloromethane/methanol 0 to 10%) to give tert-butyl N-(2-(2-((5-fluoro-4-(7-fluoro-3-cyclopentyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)amino)-7,8-dihydro-5H-1,6-naphthyridine-6-yl)-2-oxoethyl)-N-methyl-carbamate represented by formula I-22-a (100 mg, 0.158 mmol).

Step 2:

Tert-butyl N-(2-(2-((5-fluoro-4-(7-fluoro-3-cyclopentyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)amino)-7,8-dihydro-5H-1,6-naphthyridine-6-yl)-2-oxoethyl)-N-methyl-carbamate (100 mg, 0.158 mmol)(represented by formula I-22-a) was add to 2M HCl/MeOH solution (2 mL) and the mixture was stirred at room temperature for 1 hour, then concentrated to give 1-(2-((5-fluoro-4-(7-fluoro-3-cyclopentyl-2-methyl-benzimidazole-5-yl)pyrimidin-2-yl)amino)-7,8-dihydro-5H-1,6-naphthyridin-6-yl)-2-(methylamino)ethanone represented by formula I-22-b (70 mg, 0.1314 mmol).

Step 3:

1-(2-((5-Fluoro-4-(7-fluoro-3-cyclopentyl-2-methyl-benzimidazole-5-yl)pyrimidin-2-yl)amino)-7,8-dihydro-5H-1,6-naphthyridin-6-yl)-2-(methylamino)ethanone (70 mg, 0.1314 mmol) (represented by formula I-22-b), aqueous formaldehyde (30 mg) and sodium triacetoxyborohydride (128 mg) were added to 3 mL dioxane and the mixture was stirred at room temperature for 12 hours. The reaction solution was concentrated and purified by silica gel column chromatography (dichloromethane/methanol 0 to 10%) to give 2-(dimethylamino)-1-(2-((5-fluoro-4-(7-fluoro-3-cyclopentyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)amino)-7,8-dihydro-5H-1,6-naphthyridine-6-yl)ethanone represented by formula I-22 (40 mg, 0.08899 mmol). ¹H-NMR (CD₃OD) δ=8.50 (d, 1H, J=3.6 Hz), 8.23-8.27 (m, 1H), 8.17 (s, 1H), 7.77-7.80 (m, 1H), 7.55 (d, 1H, J=8.4 Hz), 4.96-5.02 (m, 2H), 4.77 (s, 1H), 4.70 (s, 1H), 3.90-3.94 (m, 2H), 2.90-3.01 (m, 2H), 2.70 (s, 3H), 2.35 (s, 4H), 2.33 (s, 2H), 2.27 (s, 4H), 2.06-2.09 (m, 2H), 1.87-1.90 (s, 2H). LC-MS: m/z (M+H)⁺=547.2.

Embodiment 23

N-(5-Fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridine-2-amine (80 mg, 0.1837 mmol)(represented by formula I-1), 4-methyl-1-piperazineacetic acid (32 mg, 0.20 mmol), HOBt (25 mg, 0.18502 mmol), EDCI (36 mg, 0.18779 mmol) were added to 1 mL DMF and the mixture was stirred at room temperature for 12 hours. The reaction solution was concentrated and purified by silica gel column chromatography (dichloromethane/methanol 0 to 10%) to give 1-(2-((5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)amino)-7,8-dihydro-5H-1,6-naphthyridin-6-yl)-2-(4-methylpiperazin-1-yl)ethanone (60 mg, 0.104 mmol). ¹H-NMR (DMSO-d₆) δ 9.99-10.02 (m, 1H), 8.69 (d, 1H, J=4 Hz), 8.30 (s, 1H), 8.11-8.16 (m, 1H), 7.70 (d, 1H, J=12 Hz), 7.55-7.72 (m, 1H), 4.81-4.89 (m, 1H), 4.75 (s, 1H), 4.60 (s, 1H), 3.77-3.88 (m, 2H), 3.23-3.25 (m, 2H), 2.90-2.95 (m, 1H), 2.65 (s, 3H), 2.33-2.43 (m, 6H), 2.16 (s, 2H), 2.09 (s, 1H), 1.64 (d, 6H, J=6.8 Hz). LC-MS: m/z (M+H)⁺=576.2.

Embodiment 26

N-(5-Fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridine-2-amine (represented by formula I-1) (80 mg, 0.1837 mmol), 3-hydroxy-3-methylbutyric acid (28 mg, 0.237 mmol), HOBt (25 mg, 0.18502 mmol), EDCI (36 mg, 0.18779 mmol) were added to 1 mL DMF and the mixture was stirred at room temperature for 12 hours. The reaction solution was concentrated and purified by silica gel column chromatography (dichloromethane/methanol 0 to 10%) to give 1-(2-((5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)amino)-7,8-dihydro-5H-1,6-naphthyridin-6-yl)-3-hydroxy-3-methyl-butanone represented by formula I-26 (10 mg, 0.019 mmol). ¹H-NMR (DMSO-d₆) δ 10.06 (d, 1H, J=11.2 Hz), 8.70 (d, 1H, J=4 Hz), 8.32 (s, 1H), 8.12-8.15 (m, 1H), 7.70 (d, 1H, J=12 Hz), 7.57-7.63 (m, 1H), 4.81-4.87 (m, 2H), 4.73 (s, 1H), 4.65 (s, 1H), 3.82-3.88 (m, 2H), 2.89-2.92 (m, 1H), 2.79-2.82 (m, 1H), 2.66 2.57-2.59 (m, 3H), 1.64 (d, 6H, J=6.8 Hz), 1.21 (s, 3H), 1.19 (s, 3H). LC-MS: m/z (M+H)⁺=536.2.

Embodiment 27

Step 1:

N-(5-Fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine (110 mg, 0.25 mmol) (represented by formula I-1), N-tert-butoxycarbonyl-2-methylalanine (100 mg, 0.49 mmol), EDCI (60 mg, 0.31 mmol), HOBt (40 mg, 0.29 mmol) and diisopropylacetamide (500 mg, 3.87 mmol) were dissolved in DMF (4 mL) and the mixture was stirred at room temperature for 16 hours. The reaction solution was concentrated and purified by silica gel column chromatography (DCMCH₃H 0 to 100%) to give tert-butyl (1-(2-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)₂-methyl-1-oxopropan-2-yl)carbamate represented by formula-27-a (70 mg) as a pale yellow solid. LC-MS m/z: (M+H)⁺=621.1.

Step 2:

Tert-butyl (1-(2-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)₂-methyl-1-oxopropan-2-yl)carbamate (70 mg, 0.11 mmol) (represented by formula I-27-a) was dissolved in 1,4-dioxane (4 mL), a solution of hydrochloride (4M) in 1,2-dioxane was added, then the mixture was stirred at room temperature for 16 hours, filtered, the cake was dissolved in water. After adjusting the pH to 7 by adding aqueous sodium bicarbonate solution, the mixture was extracted with a mixed solution of dichloromethane and methanol, and the organic layer was concentrated to give 2-amino-1-(2-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridine-6(5H)-yl)-2-methylpropyl-1-one represented by formula I-27-b (40 mg). LC-MS m/z: (M+H)⁺=521.2.

Step 3:

2-Amino-1-(2-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridine-6(5H)-yl)-2-methylpropyl-1-one (40 mg, 0.077 mmol) (represented by formula I-27-b) was dissolved in dioxane (4 mL), aqueous formaldehyde solution (10 mg, 0.33 mmol) was slowly added dropwise and the mixture was stirred at room temperature for 1 hour, then sodium triacetoxyborohydride (60 mg, 0.28 mmol) was added and stirred for 16 hours. The reaction was quenched with water, then aqueous sodium bicarbonate was added to adjust the pH to 7, the mixture was extracted with dichloromethane, the organic layer was dried over anhydrous sodium sulfate, and purified by thin-layer chromatography (DCM/CH₃OH=10/1) to give a crude product, which was dissolved in dimethyl sulfoxide, filtered, and the residue was rinsed with methanol and dried to give 2-(methylamino)-1-(2-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-hydrobenzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridine-6(5H)-yl)-2-methylpropyl-1-one (15 mg) as a yellow solid. ¹H-NMR (CDCl₃) δ 8.44 (d, 1H, J=4.0 Hz), 8.29 (d, 1H, J=6.8 Hz), 8.22 (s, 1H), 7.92-8.01 (m, 1H), 7.81 (d, 1H, J=11.6 Hz), 7.42-7.50 (m, 1H), 5.40-5.44 (m, 1H), 4.73-4.81 (m, 2H), 4.51-4.54 (m, 1H), 3.96-3.98 (m, 1H), 2.96 (t, 2H, J=4.4 Hz), 2.72 (s, 3H), 2.18-2.26 (m, 6H), 1.74 (d, 6H, J=7.2 Hz), 1.28-1.32 (m, 6H). LC-MS m/z: (M+H)⁺=548.9.

Embodiment 28

Tert-butyl 2-((5-fluoro-4-(7-fluoro-3-cyclopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)amino)-7,8-dihydro-5H-1,6-naphthyridine-6-carboxylate (7 g, 13.1 mmol) was dissolved in dioxane (40 mL) and a solution of hydrochloric acid (2M, 30 mL) in dioxane was added, the mixture was stirred at room temperature for 16 hours, concentrated, and slurried with dichloromethane to give N-(5-fluoro-4-(7-fluoro-3-cyclopropyl-2-methyl-benzimidazole-5-yl)pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amino hydrochloride represented by formula I-28 (5.5 g). ¹H-NMR (DMSO-d₆) δ 11.0 (s, 1H), 10.0 (s, 2H), 8.87 (d, 1H, J=3.2 Hz), 8.29 (s, 1H), 8.08 (d, 1H, J=8.8 Hz), 7.92-7.95 (m, 2H), 4.29 (s, 2H), 3.55-3.60 (m, 1H), 3.46-3.49 (m, 2H), 3.19 (t, 2H, J=6.0 Hz), 2.82 (s, 3H), 1.31-1.36 (m, 2H), 1.20-1.22 (m, 2H). LC-MS m/z: (M+H)⁺=434.2.

Embodiment 29

N-(5-Fluoro-4-(7-fluoro-3-cyclopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine (100 mg, 0.25 mmol) (represented by formula I-28), N,N-dimethylglycine (100 mg, 0.49 mmol), EDCI (60 mg, 0.31 mmol), HOBt (40 mg, 0.29 mmol) and diisopropylacetamide (500 mg, 3.87 mmol) were dissolved in a mixed solution of DMF (4 mL) and dichloromethane (4 mL) and the mixture was stirred at 60° C. for 16 hours. The reaction solution was concentrated and purified by silica gel column chromatography (DCM/CH₃OH/NH₃.CH₃OH=100/10/1) to give (1-(2-((4-(3-cyclopropyl-7-fluoro-2-methyl-benzimidazol-5-yl)-5-fluoro-pyrimidin-2-yl)amino-7,8-dihydro-5H-1,6-naphthyridine-6-yl)-2-(dimethylamino)-ethanone represented by formula I-29 (70 mg) as a pale yellow solid. ¹H-NMR (CDCl₃+CD₃OD) δ 8.37 (s, 1H), 8.23 (d, 1H, J=8.0 Hz), 8.11 (s, 1H), 7.72 (d, 1H, J=11.2 Hz), 7.44-7.46 (m, 1H), 4.64-4.67 (m, 2H), 3.82-3.85 (m, 2H), 3.25-3.31 (m, 4H), 2.86-2.92 (m, 2H), 2.66 (s, 3H), 2.31 (s, 6H), 1.28-1.35 (m, 2H), 1.08-1.10 (m, 2H). LC-MS m/z: (M+H)⁺=519.2.

Embodiment 30

N-(5-Fluoro-4-(7-fluoro-3-cyclopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine (100 mg, 0.25 mmol) (represented by formula I-28), N,N-morpholin-4-yl acetic acid (100 mg, 0.49 mmol), EDCI (60 mg, 0.31 mmol), HOBt (40 mg, 0.29 mmol) and diisopropylacetamide (500 mg, 3.87 mmol) were dissolved in DMF (4 mL) and the mixture was stirred at room temperature for 16 hours. The reaction solution was concentrated and purified by silica gel column chromatography (DCM/CH₃OH=100% to 90%) to give (1-(2-((4-(3-cyclopropyl-7-fluoro-2-methyl))-benzimidazol-5-yl)-5-fluoro-pyrimidin-2-yl)amino-7,8-dihydro-5H-1,6-naphthyridin-6-yl)-2-morpholine-ethanone represented by formula I-30 (20 mg) as a pale yellow solid. ¹H-NMR (CDCl₃) δ 8.44-8.46 (m, 1H), 8.30-8.33 (m, 1H), 8.16-8.18 (m, 1H), 8.09 (s, 1H), 7.78-7.82 (m, 1H), 7.50-7.52 (m, 1H), 4.74-4.79 (m, 2H), 3.92-3.95 (m, 2H), 3.75-3.77 (m, 2H), 3.67-3.72 (m, 1H), 3.31-3.35 (m, 3H), 2.94-3.05 (m, 2H), 2.76 (s, 3H), 2.55-2.57 (m, 4H), 1.33-1.38 (m, 2H), 1.13-1.17 (m, 2H). LC-MS m/z: (M+H)⁺=561.2.

Embodiment 31

3-Hydroxycyclobutylcarboxylic acid (26 mg, 0.22391 mmol), N-(5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidine-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine (80 mg, 0.1837 mmol) (represented by formula I-28), 1-hydroxybenzotriazole (25 mg, 0.185 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (36 mg, 0.18779 mmol) were added to 1 mL DMF and the mixture was stirred at room temperature for 12 hours. The reaction solution was concentrated and purified by silica gel column chromatography (dichloromethane/methanol 0 to 10%) to give (2-((5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)amino)-7,8-dihydro-5H-1,6-naphthyridin-6-yl)-(3-hydroxycyclobutyl)methanone (50 mg, 0.09370 mmol). ¹H-NMR (DMSO-d₆) δ 10.05 (d, 1H, J=12 Hz), 8.69 (d, 1H, J=3.6 Hz), 8.32 (s, 1H), 8.13 (d, 1H, J=8.4 Hz), 7.71 (d, 1H, J=12.4 Hz), 7.60 (d, 1H, J=16.4 Hz), 5.07 (s, 1H), 4.84-4.88 (m, 1H), 4.57-4.61 (m, 2H), 3.99 (s, 1H), 3.71-3.81 (m, 2H), 2.83-2.90 (m, 3H), 2.68 (s, 3H), 2.38-2.42 (m, 2H), 1.93-2.03 (m, 2H), 1.64 (d, 6H, J=6.8 Hz). LC-MS: m/z: (M+H)⁺=534.2.

Embodiment 32

3-Hydroxycyclobutylcarboxylic acid (26 mg, 0.22391 mmol), N-(5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidine-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine (80 mg, 0.1837 mmol) (represented by formula I-28), 1-hydroxybenzotriazole (25 mg, 0.185 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (36 mg, 0.18779 mmol) were added to 1 mL DMF and the mixture was stirred at room temperature for 12 hours. The reaction solution was concentrated and purified by silica gel column chromatography (dichloromethane/methanol 0 to 10%) to give (2-((5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)amino)-7,8-dihydro-5H-1,6-naphthyridin-6-yl)-(3-hydroxycyclobutyl)methanone (50 mg, 0.09370 mmol). ¹H-NMR (DMSO-d₆) δ 10.05 (d, 1H, J=12 Hz), 8.69 (d, 1H, J=3.6 Hz), 8.32 (s, 1H), 8.13 (d, 1H, J=8.4 Hz), 7.71 (d, 1H, J=12.4 Hz), 7.60 (d, 1H, J=16.4 Hz), 5.07 (s, 1H), 4.84-4.88 (m, 1H), 4.57-4.61 (m, 2H), 3.99 (s, 1H), 3.71-3.81 (m, 2H), 2.83-2.90 (m, 3H), 2.68 (s, 3H), 2.38-2.42 (m, 2H), 1.93-2.03 (m, 2H), 1.64 (d, 6H, J=6.8 Hz). LC-MS: m/z: (M+H)⁺=534.2.

Embodiment 37

N-(5-Fluoro-4-(7-fluoro-3-cyclopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine (100 mg, 0.23 mmol) (represented by formula I-28), 2-bromo-N,N-dimethylacetamide (50 mg, 0.30 mmol), diisopropylacetamide (200 mg, 1.55 mmol) were dissolved in DMF (2 mL) and the mixture was stirred at 90° C. for 16 hours. The reaction solution was cooled and diluted with water, extracted with ethyl acetate. The organic layer was concentrated and purified by silica gel thin-layer chromatography (DCM/CH₃OH=10/1). The crude product was slurried with ethyl acetate to give (2-(2-((4-(3-cyclopropyl-7-fluoro-2-methyl-benzimidazol-5-yl)-5-fluoro-pyrimidin-2-yl)amino)-7,8-dihydro-5H-1,6-naphthyridin-6-yl)-acetamide represented by formula I-37 (5 mg) as a pale yellow solid. ¹H-NMR (CDCl₃) δ 8.43 (d, 1H, J=3.6 Hz), 8.23 (d, 1H, J=8.4 Hz), 8.19 (s, 1H), 8.06 (s, 1H), 7.80 (d, 1H, J=12.0 Hz), 7.40 (d, 1H, J=8.0 Hz), 3.75 (s, 2H), 3.43 (s, 2H), 3.32-3.33 (m, 1H), 3.15 (s, 3H), 2.96-3.01 (m, 6H), 2.76 (s, 3H), 1.33-1.38 (m, 2H), 1.13-1.17 (m, 2H). LC-MS m/z: (M+H)⁺=519.2.

Embodiment 52

N-(5-Fluoro-4-(7-fluoro-3-cyclopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine (1 g, 2.3 mmol) (represented by formula I-28), 2-(dimethylamino)acetaldehyde (600 mg, 6.9 mmol) were dissolved in dichloromethane, sodium triacetoxyborohydride (1.5 g, 7.1 mmol) was slowly added, and the mixture was stirred under reflux for 16 hours. The reaction solution was concentrated and purified by silica gel column chromatography (DCM/CH₃OH/NH₃.CH₃OH=100/10/2) to give N-(4-(3-cyclopropyl-7-fluoro-2-methyl-benzimidazol-5-yl)-5-fluoro-pyrimidin-2-yl)₆-(2-dimethylaminoethyl)-7,8-dihydro-5H-1,6-naphthyridin-2-amine represented by formula I-52 (1.1 g) as a pale yellow solid. ¹H-NMR (CDCl₃) δ 8.43 (d, 1H, J=3.6 Hz), 8.26 (d, 1H, J=8.8 Hz), 8.19 (s, 1H), 7.80 (d, 1H, J=11.6 Hz), 7.42 (d, 1H, J=8.4 Hz), 3.72 (s, 2H), 3.31-3.34 (m, 1H), 3.10 (t, 2H, J=6.4 Hz), 2.92-2.98 (m, 6H), 2.76 (s, 3H), 2.74 (s, 6H), 1.33-1.38 (m, 2H), 1.13-1.17 (m, 2H). LC-MS m/z: (M+H)⁺=505.2.

Embodiment 61

N-(5-Fluoro-4-(7-fluoro-3-cyclopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine (100 mg, 0.23 mmol) (represented by formula I-28), bis(trichloromethyl)carbonate (40 mg, 0.13 mmol) and DIPEA (120 mg, 0.93 mmol) were dissolved in dichloromethane and the mixture was stirred at room temperature for 1 hour, then N,N,N′-trimethylethylenediamine (50 mg, 0.49 mmol) was added and stirred at room temperature for 16 hours. The reaction solution was concentrated, diluted with dichloromethane and water, partitioned, and the aqueous layer was concentrated and purified by column chromatography (C18, H₂O/CH₃OH=100% to 90%) and then purified by thin layer chromatography (DCM/CH₃OH=10/1) to give 2-((4-(3-cyclopropyl-7-fluoro-2-methyl-benzimidazol-5-yl)-5-fluoro-pyrimidine-)₂-yl)amino)-N-(2-dimethylaminoethyl)-N-methyl-7,8-dihydro-5H-1,6-naphthyridine-6-carboxamide represented by formula I-61 (10 mg) as a pale yellow solid. ¹H-NMR (400M Hz, CD₃OD) δ 8.45 (d, 1H, J=4.4 Hz), 7.93 (d, 1H, J=8.0 Hz), 7.79 (s, 1H), 7.43 (d, 1H, J=12.0 Hz), 7.27 (d, 1H, J=8.0 Hz), 4.28 (s, 2H), 3.52-3.55 (m, 4H), 3.24-3.27 (m, 1H), 3.04-3.07 (m, 5H), 2.80-8.83 (m, 2H), 2.72 (s, 6H), 2.58 (s, 3H), 1.22-1.24 (m, 2H), 0.95-0.98 (m, 2H). LC-MS m/z: (M+H)⁺=562.3.

Embodiment 62

Step 1:

N-(5-fluoro-4-(7-fluoro-3-cyclopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine (110 mg, 0.23 mmol) (represented by formula I-1), Boc-N-methyl-D-alanine (60 mg, 0.29 mmol), EDCI (60 mg, 0.31 mmol), HOBt (50 mg, 0.37 mmol) and diisopropylacetamide (100 mg, 0.77 mmol) were dissolved in DMF (4 mL) and the mixture was stirred at room temperature for 16 hours. The reaction solution was purified by silica gel column chromatography (DCM/CH₃OH 0 to 10%) to give tert-butyN-((1R)-(2-(2-((5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)amino)-7,8-dihydro-5H-1,6-naphthyridine-6-yl)-1-methyl-2-oxo-ethyl)-N-methylcarbamate represented by formula I-62-a (130 mg) as a pale yellow solid. LC-MS m/z: (M+H)⁺=607.2.

Step 2:

Tert-butyl N-((1R)-(2-(2-((5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)amino)-7,8-dihydro-5H-1,6-naphthyridine-6-yl)-1-methyl-2-oxo-ethyl)-N-methylcarbamate (120 mg, 0.19 mmol)(represented by formula I-62-a) was dissolved in 1,4-dioxane (4 mL), a solution of hydrochloride (4M) in 1,2-dioxane was added, the mixture was stirred at room temperature for 16 hours, filtered, dissolved in water. After adjusting the pH to 7 by adding aqueous sodium bicarbonate solution, the mixture was extracted with a mixed solution of dichloromethane and methanol, and the organic solvent was concentrated to give (2R)-1-(2-((5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6-yl)-2-(methylamino)propyl-1-one represented by the formula I-62-b (110 mg). LC-MS m/z: (M+H)⁺=507.2.

Step 3:

(2R)-1-(2-((5-Fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6-yl)-2-(methylamino)propyl-1-one (110 mg, 0.21 mmol) (represented by the formula I-62-b) was dissolved in methanol (4 mL), aqueous formaldehyde solution (10 mg, 0.33 mmol) was slowly added dropwise and the mixture was stirred at room temperature for 1 hour, then sodium cyanoborohydride (50 mg, 0.44 mmol) was added and stirred for 16 hours. The reaction solution was concentrated and purified by column chromatography (DCM/CH₃OH/NH₃.CH₃OH=10/1/0.2) to give a crude product, which was then purified by preparative liquid chromatography to give (2R)-2-(methylamino)-1-(2-((5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)amino)-7,8-dihydro-5H-1,6-naphthyridin-6-yl)propyl-1-ketone represented by formula I-62 (30 mg) as a yellow solid. ¹H-NMR (400M Hz, CD₃OD) δ 8.55 (d, 1H, J=3.6 Hz), 8.33 (d, 1H, J=1.2 Hz), 8.28 (t, 1H, J=7.2 Hz), 7.80 (d, 1H, J=12.0 Hz), 7.61-7.64 (m, 1H), 4.88-4.96 (m, 2H), 4.66-4.75 (m, 1H), 4.45-4.49 (m, 1H), 3.85-4.11 (m, 2H), 3.03-3.11 (m, 1H), 2.95-2.98 (m, 1H), 2.84 (d, 6H, J=3.6 Hz), 2.71 (s, 3H), 1.74 (d, 6H, J=6.8 Hz), 1.50-1.56 (m, 3H). LC-MS m/z: (M+H)⁺=535.2.

Embodiment 72

Step 1:

N-(5-Fluoro-4-(7-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-5-yl)pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine (200 mg, 0.46 mmol) (represented by formula I-1) was dissolved in 5 mL methanol, 1-tert-butoxycarbonyl-3-pyrrolidone (170 mg, 0.92 mmol) and sodium triacetoxyborohydride (146 mg, 0.92 mmol) were added, the mixture was stirred at room temperature for 16 hours. Solvent was evaporated under reduced pressure, saturated sodium bicarbonate solution was added, the mixture was extracted with dichloromethane, evaporated under reduced pressure. The residue was purified by silica gel column chromatography to give the compound represented by formula I-72-a as a yellow solid (170 mg), yield 61%. LC-MS: 604.9[M+H]⁺.

Step 2:

Tert-butyl 3-(2-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridine-6(5H)-yl)pyrrolidine-1-carboxylate (represented by formula I-72-a) (170 mg, 0.28 mmol) was dissolved in 5 mL dioxane, a solution of 4N HCl in dioxane (1 mL) was added dropwise, and the mixture was stirred at room temperature for 4 hours. Solvent was evaporated under reduced pressure, saturated sodium bicarbonate solution was added, and the mixture was extracted with dichloromethane. The solvent was evaporated under reduced pressure to give the compound represented by the formula I-72-b as a yellow solid (138 mg), yield 97%. LC-MS: 504.9 [M+H]⁺.

Step 3:

N-(5-Fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)-6-(pyrrolidine-3-yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine (represented by formula I-72-b) (138 mg, 0.27 mmol) was dissolved in 5 mL dioxane, sodium hydroxide (55 mg, 1.37 mmol) and bromopropyne (32 mg, 0.27 mmol) were added and the mixture was stirred at room temperature for 2 hours. Solvent was evaporated under reduced pressure, and the residue was purified by silica gel column chromatography to give the compound represented by formula I-72 as a yellow solid, 55 mg, 38% yield. ¹H NMR (400 MHz, CDCl₃) δ 8.43 (d, J=3.8 Hz, 1H), 8.23 (dd, J=9.2, 4.8 Hz, 2H), 7.99 (s, 1H), 7.80 (d, J=12.2 Hz, 1H), 7.39 (d, J=8.5 Hz, 1H), 4.75 (dt, J=14.0, 7.0 Hz, 1H), 3.69 (s, 2H), 3.49 (d, J=2.2 Hz, 2H), 3.27-3.05 (m, 2H), 3.00 (d, J=7.9 Hz, 2H), 2.97-2.75 (m, 4H), 2.74-2.68 (m, 3H), 2.28 (t, J=2.3 Hz, 1H), 2.20 (dd, J=13.0, 6.3 Hz, 1H), 1.92 (dd, J=12.9, 6.4 Hz, 2H), 1.73 (d, J=7.0 Hz, 6H). LC-MS: m/z: (M+H)⁺=542.9.

Embodiment 73

Step 1:

N-(5-Fluoro-4-(7-fluoro-3-cyclopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine (100 mg, 0.23 mmol) (represented by formula I-28), 2(R)-2-chloromethyloxirane (30 mg, 0.32 mmol) and DIPEA (100 mg, 0.77 mmol) were dissolved in N,N-dimethylformamide (4 mL), the mixture was stirred at 80° C. for 16 hours. The reaction solution was concentrated and purified by silica gel column chromatography (DCM/CH₃OH=10/1) to give (S)—N-(4-(1-cyclopropyl-4-fluoro-2-methyl-1H-benzimidazol-6-yl)-5-fluoropyrimidin-2-yl)-6-(epoxy-2-ylmethyl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine represented by formula I-73-a (30 mg) as a pale yellow solid. LC-MS m/z: (M+H)⁺=490.2.

Step 2:

(S)—N-(4-(1-cyclopropyl-4-fluoro-2-methyl-1H-benzimidazol-6-yl)-5-fluoropyrimidin-2-yl)-6-(epoxy-2-ylmethyl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine (30 mg, 0.06 mmol)(represented by formula I-73-a), dimethylamine (10 mg, 0.22 mmol) were dissolved in ethanol (5 mL), the mixture was stirred at 60° C. for 4 hours. The reaction solution was concentrated and purified by silica gel column chromatography (DCM/CH₃OH=10/1) to give (S)-1-(2-((4-(1-cyclopropyl-4-fluoro-2-methyl-1H-benzimidazol-6-yl)-5-fluoropyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridine-6(5H)-yl)-3-(dimethylamino)propyl-2-ol represented by formula I-73 (10 mg) as a pale yellow solid. H NMR (400 MHz, CDCl₃) δ 8.37 (d, J=3.7 Hz, 1H), 8.19 (d, J=8.5 Hz, 1H), 8.13 (s, 1H), 7.77-7.70 (m, 1H), 7.40 (d, J=8.6 Hz, 1H), 4.35 (m, 1H), 3.77 (s, 2H), 3.23 (d, J=15.2 Hz, 2H), 3.06-2.91 (m, 5H), 2.87 (s, 6H), 2.72-2.62 (m, 5H), 1.34-1.29 (m, 2H), 1.10 (t, J=8.0 Hz, 2H). LC-MS m/z: (M+H)⁺=534.9.

Embodiment 77

Step 1:

N-(5-Fluoro-4-(7-fluoro-3-cyclopropyl-2-methyl-benzoimidazol-5-yl)pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine (100 mg, 0.23 mmol) (represented by formula I-28), N-boc-D-alanal (100 mg, 0.58 mmol) were dissolved in dichloromethane, sodium triacetoxyborohydride (200 mg, 0.94 mmol) was added slowly, the mixture was stirred under reflux for 16 hours. The reaction solution was concentrated and purified by silica gel column chromatography (DCM/CH₃OH/NH₃.CH₃OH=100/10/2) to give N-(4-(3-tert-butyl-N-((1R)-2-(2-((4-(3-cyclopropyl-7-fluoro-2-methyl-benzoimidazol-5-yl)-5-fluoro-pyrimidin-2-yl)amino)-7,8-dihydro-5H-1,6-naphthyridin-6-yl)-1-methyl-ethyl)carbonate represented by formula I-77-a (60 mg) as a pale yellow solid. LC-MS m/z: (M+H)⁺=591.2.

Step 2:

Tert-butyl N-((1R)-2-(2-((4-(3-cyclopropyl-7-fluoro-2-methyl-benzimidazol-5-yl)-5-fluoro-pyrimidine2-yl)amino)-7,8-dihydro-5H-1,6-naphthyridin-6-yl)-1-methyl-ethyl)carbonate (60 mg, 0.2 mmol) (represented by formula I-77-a) was dissolved in dichloromethane (4 mL), a solution of hydrochloric acid in 1,2-dioxane (4M, 1 mL) was added, and the mixture was stirred at room temperature for 4 hours. The reaction solution was concentrated and dissolved in dichloromethane and stirred at room temperature for 30 min, then filtered and dried to give 6-((2R)-aminopropyl)-N-(4-(3-cyclopropyl-7-fluoro-2-yl-benzimidazol-5-yl)-5-fluoro-pyrimidin-2-yl)-7,8-dihydro-5H-1,6-naphthyridin-2-amino hydrochloride (60 mg) as a pale yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 8.98 (d, J=3.3 Hz, 1H), 8.58 (s, 1H), 8.27 (d, J=11.2 Hz, 1H), 8.22 (d, J=9.1 Hz, 1H), 7.59 (d, J=9.0 Hz, 1H), 4.75-4.44 (m, 2H), 4.08 (M, 1H), 3.95-3.82 (m, 1H), 3.80-3.73 (m, 2H), 3.67-3.48 (m, 4H), 3.04 (s, 3H), 1.54 (M, 5H), 1.42-1.34 (m, 2H). LC-MS: m/z: (M+H)⁺=490.9.

Embodiment 80

6-((2R)-Aminopropyl)-N-(4-(3-cyclopropyl-7-fluoro-2-methyl-benzimidazol-5-yl)-5-fluoro-pyrimidine-2-yl)-7,8-dihydro-5H-1,6-naphthyridin-2-amine (50 mg, 0.21 mmol) (represented by formula I-77) was dissolved in methanol (6 mL), aqueous formaldehyde (20 mg, 0.67 mmol) was slowly added dropwise, followed by addition of sodium triacetoxyborohydride (50 mg, 0.24 mmol), the mixture was stirred at 60° C. for 16 hours. The reaction solution was concentrated and purified by column chromatography (DCM/CH₃OH/NH₃.CH₃OH=10/1/0.2) to give a crude product, which was purified by preparative liquid phase to give N-(4-(3-cyclopropyl-7-fluoro-2-methyl-benzoimidazol-5-yl)-5-fluoro-pyrimidin-2-yl)-6-((2R)-2-(dimethylaminopropyl)-7,8-dihydro-5H-1,6-naphthyridin-2-amine represented by formula I-80 (25 mg) as a pale yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 8.52 (d, J=3.9 Hz, 1H), 8.26 (s, 1H), 8.18 (d, J=8.5 Hz, 1H), 7.80 (d, J=12.1 Hz, 1H), 7.52 (d, J=8.6 Hz, 1H), 3.87 (d, J=14.7 Hz, 1H), 3.78-3.65 (m, 2H), 3.46 (m, 1H), 3.14-3.04 (m, 1H), 3.02-2.88 (m, 4H), 2.80 (s, 6H), 2.75 (s, 3H), 2.68 (m, 1H), 1.36 (d, J=7.0 Hz, 2H), 1.31 (d, J=6.6 Hz, 3H), 1.21-1.14 (m, 2H). LC-MS: m/z: (M+H)⁺=519.0

Embodiment 95

Step 1:

Potassium carbonate (75 mg, 0.55 mmol) and N-(5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidine-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine (120 mg, 0.275 mmol) was added to 15 mL N,N-dimethylformamide, followed by the addition of ethyl bromoacetate (75 mg, 0.45 mmol), the mixture was stirred at room temperature overnight. The reaction solution was evaporated to dryness under reduced pressure and purified by silica gel column chromatography to give 2-(2-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethyl acetate represented by the formula I-95-a (100 mg) as a pale yellow solid.

Step 2:

Ethyl 2-(2-((5-Fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl) acetate was dissolved in 15 mL dry dichloromethane, and 2 mL DIBAL-H (1 mol/L in toluene) was added at −78° C., the mixture was stirred at this temperature for 3h. Then the reaction was quenched with methanol, 2 mL saturated ammonium chloride was added and the mixture was stirred at room temperature for 10 minutes. The organic layer was dried over sodium sulfate and then concentrated to give 2-(2-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)acetaldehyde represented by formula I-95-b, which was used in the next step without further purification. LC-MS: m/z: (M+H)⁺=478.

Step 3:

2-(2-((5-Fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)acetaldehyde (80 mg, 0.167 mmol), sodium acetate (100 mg, 0.73 mmol) and hydroxylamine hydrochloride (70 mg, 1 mmol) were added to 15 mL ethanol and the mixture was stirred at 80° C. for 2 h. The reaction solution was evaporated to dryness under reduced pressure and then purified by silica gel column chromatography to give 2-(2-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)acetaldehyde represented by formula I-95 (28 mg) as a light yellow solid. H NMR (400 MHz, CD₃OD) δ 8.48 (d, J=3.8 Hz, 1H), 8.28 (d, J=0.9 Hz, 1H), 8.22 (d, J=8.5 Hz, 1H), 7.78 (d, J=11.9 Hz, 1H), 7.55-7.42 (m, 1.44H), 6.90 (t, J=4.5 Hz, 0.3H), 4.89-4.83 (m, 1H), 3.71 (d, J=10.8 Hz, 2H), 3.58 (d, J=4.5 Hz, 1H), 3.36 (d, J=6.1 Hz, 2H), 3.06-2.89 (m, 4H), 2.69 (s, 3H), 1.74 (t, J=6.5 Hz, 6H). LC-MS: m/z: (M+H)⁺=493.

Embodiment 97

Step 1:

Tert-butyl-N—(N-tert-butoxycarbonyl-N′-(trifluoromethylsulfonyl)carbamimidoyl)carbamate (80 mg, 0.21 mmol), 6-(2-aminoethyl)-N-(5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine hydrochloride (100 mg, 0.21 mol) and triethylamine (0.5 ml) were added to 15 mL N,N-dimethylformamide and the mixture was stirred at room temperature overnight. The reaction solution was evaporated to dryness under reduced pressure and then purified by silica gel column chromatography to give tert-butyl-N—(N-tert-butoxycarbonyl-N′-(2-(2-((5-fluoro-4-(7-fluoro-3-isopropyl-2-methylbenzimidazol-5-yl)pyrimidin-2-yl)amino)-7,8-dihydro-5H-1,6-naphthyridin-6-yl)ethyl)carbamimidoyl)carbamic acid represented by the formula I-97-a (130 mg) as a pale yellow solid. LC-MS: m/z: (M+H)⁺=721.

Step 2:

Tert-butyl-N—(N-tert-butoxycarbonyl-N′-(2-(2-((5-fluoro-4-(7-fluoro-3-isopropyl-2-methylbenzimidazol-5-yl)pyrimidin-2-yl)amino)-7,8-dihydro-5H-1,6-naphthyridin-6-yl)ethyl)carbamimidoyl)carbamic acid (120 mg, 0.166 mmol) was added to 5 mL dichloromethane and 5 mL trifluoroacetic acid was added under an ice bath. The mixture was stirred at room temperature for 3h, then evaporated to dryness under reduced pressure and purified by preparative HPLC to give 2-(2-(2-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridine-6(5H)-yl)ethyl)guanidine represented by formula I-97 (50 mg) as a light yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 8.53 (d, J=3.9 Hz, 1H), 8.32 (d, J=1.2 Hz, 1H), 8.26 (s, 1H), 8.16 (d, J=8.5 Hz, 1H), 7.77 (d, J=12.8 Hz, 1H), 7.50 (d, J=8.6 Hz, 1H), 4.93 (d, J=6.9 Hz, 1H), 3.73 (s, 2H), 3.48-3.41 (m, 2H), 2.97 (s, 4H), 2.84 (t, J=5.7 Hz, 2H), 2.71 (s, 3H), 1.73 (d, J=6.9 Hz, 6H). LC-MS: m/z: (M+H)⁺=521.

Embodiment 107

Triethylamine (0.5 mL) and N-(5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidine-2-yl)-6-(2-(methylamino)ethyl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine hydrochloride (100 mg, 0.203 mmol) were added to 15 mL dry dichloromethane, then dimethylphosphine chloride (30 mg, 0.2667 mmol) was added and the mixture was stirred at room temperature for 3 h. The reaction solution was evaporated to dryness under reduced pressure and purified by silica gel column chromatography (DCM/CH₃OH/NH₃.CH₃OH=10/1/0.2) to give N-(2-(2-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethyl)-N,P,P-dimethylphosphinic amide represented by formula I-107 (40 mg) as a pale yellow solid. H NMR (400 MHz, CDCl₃) δ 8.44 (d, J=3.8 Hz, 1H), 8.22 (dd, J=17.5, 9.9 Hz, 3H), 7.78 (d, J=11.6 Hz, 1H), 7.39 (d, J=8.5 Hz, 1H), 4.83-4.67 (m, 1H), 3.77 (s, 2H), 3.29 (dd, J=15.9, 7.0 Hz, 2H), 3.00 (s, 4H), 2.83-2.77 (m, 2H), 2.72 (d, J=11.8 Hz, 6H), 1.74 (t, J=9.7 Hz, 6H), 1.50 (d, J=13.2 Hz, 6H). LC-MS: m/z: (M+H)⁺=569.

Embodiment 110

N-(5-Fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)-6-(2-(methylamino)ethyl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine hydrochloride (100 mg, 0.2 mmol), diisopropylethylamine (0.5 mL) and thiocarbonyldiimidazole (74 mg, 0.41 mmol) were added to 15 mL N,N-dimethylformamide, the mixture was stirred at 50° C. for 3h. The reaction solution was cooled to room temperature, 10 mL aqueous ammonia was added to the above reaction solution, and the mixture was stirred overnight at room temperature. The reaction solution was evaporated to dryness under reduced pressure, then purified by silica gel column chromatography to give 1-(2-(2-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridine-6(5H)-yl)ethyl)-1-methylthiourea represented by formula I-110 (24 mg) as a pale yellow solid. ¹H NMR (400 MHz, DMSO-d₆) δ 9.95 (s, 1H), 8.68 (d, J=3.9 Hz, 1H), 8.32 (s, 1H), 8.07 (d, J=8.5 Hz, 1H), 7.70 (d, J=12.0 Hz, 1H), 7.45 (d, J=8.5 Hz, 1H), 7.25 (s, 1H), 4.85 (dt, J=13.9, 7.0 Hz, 1H), 3.64 (s, 2H), 3.11 (d, J=19.9 Hz, 2H), 2.86 (d, J=11.9 Hz, 4H), 2.72 (t, J=6.5 Hz, 2H), 2.65 (s, 3H), 1.64 (d, J=6.9 Hz, 6H). LC-MS: m/z: (M+H)⁺=552.

Embodiment 132

6-(2-Aminoethyl)-N-(5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidine-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine hydrochloride (120 mg, 0.25 mol) and 1H-pyrazole-1-carboxamidine hydrochloride (73 mg, 0.5 mmol) were dissolved in N,N-dimethylformamide (5 ml), then diisopropylethylamine (350 mg, 2.7 mmol) was added and the mixture was stirred at room temperature overnight. The reaction solution was evaporated under reduced pressure, and then purified by preparative HPLC to give 1-(2-(2-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridine-6(5H)-yl)ethyl)guanidine (28 mg) as a pale yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 8.51 (d, J=4.0 Hz, 1H), 8.38 (s, 2H), 8.29 (d, J=1.1 Hz, 1H), 8.13 (d, J=8.5 Hz, 1H), 7.71 (d, J=12.7 Hz, 1H), 7.44 (d, J=8.5 Hz, 1H), 4.93 (d, J=7.0 Hz, 0H), 3.69 (s, OH), 3.45 (t, J=5.6 Hz, 2H), 2.93 (s, 4H), 2.82 (t, J=5.6 Hz, 2H), 2.69 (s, 3H), 1.72 (d, J=6.9 Hz, 6H). LC-MS: m/z: (M+H)⁺=521.

Embodiment 133

Step 1:

N-(5-Fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)-6-(2-(methylamino)ethyl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine hydrochloride (90 mg, 0.15 mmol), tert-butyl (2-oxoethyl)carbamic acid (50 mg, 0.31 mmol) and triethylamine (1 mL) were added to 25 mL dichloromethane, followed by the addition of sodium triacetoxyborohydride (65 mg, 0.3 mmol). The mixture was stirred at room temperature overnight. The reaction solution was evaporated to dryness under reduced pressure and the residue was purified by silica gel column chromatography to give tert-butyl(2-((2-(2-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridine-6(5H)-yl)ethyl)(methyl)amino)ethyl)carbamate represented by formula I-133-a (80 mg) as a pale yellow solid. LC-MS: m/z: (M+H)⁺=636.

Step 2:

Tert-butyl(2-((2-(2-((5-Fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridine-6(5H)-yl)ethyl)(methyl)amino)ethyl)carbamate (90 mg, 0.14 mmol) was dissolved in methanol (3 mL), 4 mL solution of 4 mol/L hydrochloride in 1,4-dioxane was added, and the resulting solution was stirred at room temperature for 2h. The reaction mixture was evaporated to dryness under reduced pressure to give N¹-(2-(2-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridine-6(5H)-yl)ethyl)-N¹-methylethane-1,2-diamine hydrochloride represented by formula I-133-b (80 mg) as a white solid. LC-MS: m/z: (M+H)⁺=536.

Step 3:

N¹-(2-(2-((5-Fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridine-6(5H)-yl)ethyl)-NI-methylethane-1,2-diamine hydrochloride (80 mg, 0.15 mmol) and 1H-pyrazole-1-carboxamidine hydrochloride (45 mg, 0.3 mmol) were dissolved in N,N-dimethylformamide (5 mL) and diisopropylethylamine (350 mg, 2.7 mmol) was added. The mixture was stirred at room temperature overnight, then evaporated to dryness under reduced pressure and purified by preparative HPLC to give 1-(2-((2-(2-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridine-6(5H)-yl)ethyl)(methyl)amino)ethyl)guanidine (29 mg) as a pale yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 8.54 (d, J=3.9 Hz, 1H), 8.40 (s, 1H), 8.31 (d, J=1.0 Hz, 1H), 8.22 (d, J=8.5 Hz, 1H), 7.78 (d, J=12.0 Hz, 1H), 7.54 (d, J=8.6 Hz, 1H), 4.93 (d, J=6.9 Hz, 1H), 3.97 (s, 2H), 3.37 (t, J=5.7 Hz, 2H), 3.21 (t, J=6.0 Hz, 2H), 3.07 (t, J=5.7 Hz, 2H), 3.02 (t, J=6.2 Hz, 2H), 2.86 (t, J=6.1 Hz, 2H), 2.77 (t, J=5.7 Hz, 2H), 2.71 (s, 3H), 2.42 (s, 3H), 1.73 (d, J=6.9 Hz, 6H). LC-MS: m/z: (M+H)⁺=578.

Embodiment 135

N-(5-Fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)-6-(2-(methylamino)ethyl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine hydrochloride (100 mg, 0.2 mmol), diisopropylethylamine (0.5 mL) and thiocarbonyldiimidazole (74 mg, 0.41 mmol) were added to 15 mL N,N-dimethylformamide and the mixture was stirred at 50° C. for 3 hours. The reaction mixture was cooled to room temperature and 30 mL methanol was added. The mixture was stirred overnight at room temperature, then evaporated to dryness under reduced pressure, and purified by silica gel column chromatography to give N-(2-(2-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridine-6(5H)-yl)ethyl)-N-methyl-1H-imidazole-1-carbothioamide represented by formula I-135 (21 mg) as a pale yellow solid. ¹H NMR (400 MHz, DMSO-d₆) δ 10.09 (s, 1H), 8.71 (d, J=3.9 Hz, 1H), 8.31 (s, 1H), 8.16 (d, J=8.6 Hz, 1H), 7.88 (s, 1H), 7.70 (d, J=12.2 Hz, 1H), 7.45 (d, J=8.3 Hz, 1H), 7.23 (s, 1H), 7.00 (s, 1H), 5.37 (s, 2H), 4.90-4.76 (m, 1H), 3.89-3.75 (m, 2H), 3.55 (d, J=3.7 Hz, 4H), 3.05-2.95 (m, 5H), 2.65 (s, 3H), 1.60 (d, J=6.9 Hz, 6H). LC-MS: m/z: (M+H)⁺=603.

Embodiment 137

Step 1:

Diisopropylethylamine (0.5 mL) and N-(5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine (132 mg, 0.3 mmol) were added to 15 mL N,N-dimethylformamide, bromoacetonitrile (72 mg, 0.6 mmol) was added and the mixture was stirred at room temperature for 2h. The reaction solution was evaporated to dryness under reduced pressure and then purified by silica gel column chromatography to give 2-(2-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)acetonitrile represented by formula I-137-a (120 mg) as a pale yellow solid. LC-MS: m/z: (M+H)⁺=475.

Step 2:

2-(2-((5-Fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)acetonitrile (120 mg, 0.253 mmol) was added to a mixed solvent of 20 mL ethanol and 20 mL tetrahydrofuran, then 0.5 mL 50% aqueous hydroxylamine solution was added and the mixture was stirred at 80° C. overnight. The reaction solution was evaporated to dryness under reduced pressure and then purified by silica gel column chromatography to give 2-(2-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridine-6(5H)-yl)-N′-hydroxyacetimidamide represented by formula I-137 (85 mg) as a pale yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 8.47 (d, J=3.9 Hz, 1H), 8.27 (d, J=0.9 Hz, 1H), 8.19 (d, J=8.5 Hz, 1H), 7.75 (d, J=11.5 Hz, 1H), 7.43 (d, J=8.5 Hz, 1H), 4.87 (d, J=7.0 Hz, 1H), 3.64 (s, 2H), 3.20 (s, 2H), 2.96 (d, J=5.2 Hz, 2H), 2.88 (t, J=5.6 Hz, 2H), 2.69 (s, 3H), 1.73 (d, J=6.9 Hz, 6H). LC-MS: m/z: (M+H)⁺=508.

Embodiment 153

N-(4-(3-Tert-butyl-7-fluoro-2-methyl-benzimidazol-5-yl)-5-fluoro-pyrimidine-2-yl)-6-(2-chloroethyl)-7,8-dihydro-5H-1,6-naphthyridin-2-amine (50 mg, 0.09766 mmol) (represented by formula 6-b), anhydrous acetonitrile (1 mL) and morpholine (85 mg, 0.9766 mmol) were successively added to a dry flask at room temperature under argon atmosphere. The reaction mixture was stirred at 80° C. for 12 hours. LCMS monitored the reaction was complete. The reaction mixture was cooled to room temperature and concentrated by a rotary evaporator. The residue was purified by preparative TLC (silica gel, dichloromethane/methanol=10:1, v/v) to give N-(4-(3-tert-butyl-7-fluoro-2-methyl-benzimidazol-5-yl)-5-fluoro-pyrimidin-2-yl)-6-(2-diethylaminoethyl)-7,8-dihydro-5H-1,6-naphthyridin-2-amine represented by formula I-153 (12.8 mg, 0.0227 mmol) as a pale yellow solid. ¹H NMR (400 MHz, Chloroform-d) δ 8.41 (d, J=3.8 Hz, 1H), 8.38 (d, J=1.2 Hz, 1H), 8.32 (br.s, 1H), 8.24 (d, J=8.5 Hz, 1H), 7.74 (d, J=11.3 Hz, 1H), 7.35 (d, J=8.5 Hz, 1H), 3.84-3.68 (m, 6H), 3.02-2.93 (m, 4H), 2.87 (s, 3H), 2.85-2.79 (m, 2H), 2.75-2.69 (m, 2H), 2.66-2.59 (m, 4H), 1.90 (s, 9H). LC-MS m/z:(M+H)⁺=563.40.

Embodiment 155

Step 1:

Vinylmagnesium bromide (25 mL, 0.7 mmol/L) was added to a solution of methylphosphonic dichloride (1 g, 7.52 mmol) in tetrahydrofuran at −78° C. in more than half an hour. The mixture was stirred at this temperature for 3.5 hours, then warmed to 0° C. and stirred for 1 hour. The reaction was quenched with 20 mL saturated ammonium chloride solution. The mixture was filtered through a pad of silica gel and eluted with 10% 7M methanolic ammonia in dichloromethane. The filtrate was concentrated to give a crude product (0.1 g) as colorless viscous liquid, yield 10%, which was directly used in the next step.

Step 2:

Methyl divinyl phosphine oxide (20 mg, 0.17 mmol) and 6-(2-aminoethyl)-N-(4-(1-(tert-butyl)-4-fluoro-2-methyl-1H-benzo[d]imidazol-6-yl)-5-fluoropyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine (50 mg, 0.1 mmol) were added to a mixed solvent of tetrahydrofuran (6 mL) and water (2 mL), then 0.5 mL triethylamine was added and the mixture was stirred under reflux overnight. The reaction mixture was evaporated to dryness under reduced pressure and purified by column chromatography (methanol:dichloromethane=0-20%) to give a crude product, which was purified by preparative HPLC to give 1-(2-(2-((4-(1-(tert-butyl)-4-fluoro-2-methyl-1H-benzo[d]imidazol-6-yl)-5-fluoropyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethyl)-4-methyl-1,4-azaphosphinane 4-oxide (8 mg) as a yellow solid, yield 12%. ¹H NMR (400 MHz, CDCl₃) δ 8.43 (d, J=8.2 Hz, 2H), 8.26 (d, J=8.1 Hz, 1H), 7.77 (d, J=10.9 Hz, 1H), 7.42 (d, J=8.6 Hz, 1H), 3.74 (s, 2H), 2.98 (s, 6H), 2.83 (m, 9H), 1.95 (t, 13H), 1.57 (d, J=12.9 Hz, 3H). LC-MS m/z: (M+H)⁺=609.

Embodiment 187

Step 1:

N-(5-Fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine (400 mg, 0.918 mmol) and diisopropylethylamine (ml) were added to a mixed solvent of 10 mL N,N-dimethylformamide and 20 mL acetonitrile, then 2-chloroethyl methyl sulfide (0.6 mL) was added and the mixture was stirred at 70° C. overnight. Additional 2-chloroethyl methylsulfide (0.6 mL) was added and stirred at 70° C. overnight. The mixture was concentrated under reduced pressure and purified by silica gel column chromatography to give N-(5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)-6-(2-(methylthio)ethyl)-5,6,7,8-tetrahydro-1,6-naphthyridine-2-amine (210 mg) as a yellow solid. LC-MS: m/z: (M+H)⁺=510.

Step 2:

N-(5-Fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)-6-(2-(methylthio)ethyl)-5,6,7,8-tetrahydro-1,6-naphthyridine-2-amine (100 mg, 0.1962 mmol) was added to 10 mL dichloromethane, then m-chloroperbenzoic acid (42 mg, 0.188 mmol) was added and the mixture stirred overnight at room temperature. The mixture was concentrated under reduced pressure and purified by silica gel column chromatography to give N-(5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)-6-(2-(methylsulfonyl)ethyl)-5,6,7,8-tetrahydro-1,6-naphthyridine-2-amine represented by the formula I-187-b (80 mg) as a pale yellow solid. LC-MS: m/z: (M+H)⁺=526.

Step 3:

N-(5-Fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)-6-(2-(methylsulfonyl)ethyl)-5,6,7,8-tetrahydro-1,6-naphthyridine-2-amine (90 mg, 0.17 mmol) and 2-((aminooxy)sulfonyl)-1,3,5-trimethylbenzene (60 mg, 0.28 mmol) were added to a mixed solvent of 10 mL dichloromethane and 10 mL acetonitrile, the mixture was stirred at room temperature overnight. The solvent was evaporated under reduced pressure and the crude product was purified by preparative HPLC to give N-(5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazol-5-yl)pyrimidin-2-yl)-6-(2-(methylsulfonyl)ethyl]-7,8-dihydro-5H-1,6-naphthyridine-2-amine 2,4,6-trimethylbenzenesulfonate represented by formula I-187 (30 mg) as a pale yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 8.56 (d, J=3.8 Hz, 1H), 8.40-8.29 (m, 2H), 7.79 (d, J=11.8 Hz, 1H), 7.60 (d, J=8.8 Hz, 1H), 6.88 (s, 2H), 4.92 (d, J=6.9 Hz, 2H), 4.78 (s, 1H), 4.07 (s, 4H), 3.76-3.61 (m, 1H), 3.52-3.39 (m, 2H), 3.28 (dd, J=14.5, 9.5 Hz, 1H), 2.82 (d, J=3.3 Hz, 3H), 2.71 (s, 3H), 2.63 (s, 6H), 2.24 (s, 3H), 1.73 (d, J=6.9 Hz, 6H). LC-MS: m/z: (M+H)⁺=541.

Embodiment 197

Diethyl(2-((2-(2-((4-(1-(tert-butyl)-4-fluoro-2-methyl-1H-benzo[d]imidazol-6-yl)-5-fluoropyrimidin-2(5H)-yl)ethyl)(methyl)amino)ethyl)phosphonate (100 mg, 0.15 mmol) was added to 10 mL dichloromethane, followed by the addition of trimethylsilyl iodide (0.2 mL). The mixture was stirred at room temperature overnight. The reaction mixture was evaporated to dryness under reduced pressure and the crude product was purified by preparative HPLC to give (2-((2-(2-((4-(1-(tert-butyl)-4-fluoro-2-methyl-1H-benzo[d]imidazol-6-yl)-5-fluoropyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethyl)(methyl)amino)ethyl)phosphonic acid (50 mg) as a pale yellow solid, yield 54%. ¹H NMR (400 MHz, MeOD) δ 8.47 (d, J=3.8 Hz, 1H), 8.41 (d, J=1.1 Hz, 1H), 8.17 (d, J=8.6 Hz, 1H), 8.08 (s, 1H), 7.75 (d, J=11.5 Hz, 1H), 7.51 (d, J=8.6 Hz, 1H), 3.87 (s, 2H), 3.24 (m, 4H), 3.13-3.00 (m, 6H), 2.85 (s, 3H), 2.80 (s, 3H), 2.04-1.82 (m, 12H). LC-MS: m/z: (M+H)⁺=615.

Embodiment 204

N-(4-(3-cyclopropyl-7-fluoro-2-methyl-benzimidazol-5-yl)-5-fluoro-pyrimidin-2-yl)-6-(2-dimethylaminoethyl)-7,8-dihydro-5H-1,6-naphthyridin-2-amine (100 mg, 0.198 mmol) was dissolved in hydrogen peroxide (5 mL), the mixture was stirred at room temperature for about 1 hour, then quenched by adding aqueous sodium sulfite solution. Water was distilled off under reduced pressure. The solid was dissolved in a mixed solvent of dichloromethane and methanol, then filtered and the filtrate was purified by HPLC to give 2-(2-((4-(3-cyclopropyl-7-fluoro-2-methyl-benzimidazol-5-yl)-5-fluoro-pyrimidin-2-yl)amino)₆-oxo-7,8-dihydro-5H-1,6-naphthyridine-6-yl)-N,N-dimethyl-ethylaminooxy represented by formula I-205 (30 mg) as a pale yellow solid. ¹H NMR (400 MHz, MeOD) δ 8.61-8.47 (br, 2H), 8.02 (s, 1H), 7.52-7.30 (br, 2H), 4.60-4.40 (br, 4H), 4.14-3.85 (m, 2H), 3.56-3.70 (br, 4H), 3.48-3.35 (m, 2H), 3.02-2.85 (m, 1H), 2.55 (s, 3H), 1.30-1.15 (m, 2H), 1.05-0.84 (m, 2H). LC-MS: m/z: (M+H)⁺=537.

Embodiment 205

Diethyl phosphonate (50 mg, 0.36 mmol), triethylamine (0.5 ml) and N-(5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)-6-(2-(methylamino)ethyl)-5,6,7,8-tetrahydro-1,6-naphthyridine-2-amine (120 mg, 0.24 mmol) were added to a mixed solvent of 10 mL acetonitrile, 10 mL carbon tetrachloride and 5 mL dichloromethane, and the mixture was stirred at room temperature overnight. The reaction mixture was filtered and the filtrate was evaporated to dryness under reduced pressure to give a crude product, which was purified by preparative HPLC to give (2-(2-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidine-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethyl)(methyl)phosphoramidate (90 mg) as a pale yellow solid, yield 58%. ¹H NMR (400 MHz, CDCl₃) δ 8.44 (d, J=3.8 Hz, 1H), 8.28 (d, J=8.5 Hz, 1H), 8.23 (d, J=1.1 Hz, 1H), 7.80 (d, J=11.7 Hz, 1H), 7.42 (d, J=8.6 Hz, 1H), 4.76 (dd, J=13.9, 6.9 Hz, 1H), 4.10-3.99 (m, 4H), 3.94 (dt, J=14.2, 6.8 Hz, 2H), 3.83 (s, 2H), 3.38 (m, 2H), 3.05 (s, 3H), 2.86 (t, J=6.9 Hz, 2H), 2.76-2.69 (m, 5H), 1.74 (d, J=7.1 Hz, 6H), 1.37-1.27 (t, 6H).

LC-MS m/z: (M+H)⁺=607.

TABLE 1 List of embodiments Embodi- ment Structure Analytic data Method 24

White powder. ¹HNMR (400 MHz, DMSO-d₆) δ 10.05 (d, J = 6.7 Hz, 1H), 8.69 (s, 1H), 8.31 (s, 1H), 8.14 (t, J = 11.4 Hz, 1H), 7.71 (d, J = 11.9 Hz, 1H), 7.65-7.56 (m, 1H), 4.86 (t, J = 5.4 Hz, 1H), 4.68 (d, J = 40.6 Hz, 2H), 3.90- 3.80 (m, 2H), 3.62 (d, J = 18.6 Hz, 4H), 3.42 (d, J = 40.8 Hz, 4H), 2.88 (d, J = 55.2 Hz, 3H), 2.66 (s, 4H), 1.65 (d, J = 6.9 Hz, 7H). LC-MS: m/z: (M + H)⁺ = 563.2. I-5 25

Yellow powder. ¹H NMR (400 MHz, CD₃OD) δ 8.47 (d, J = 51.2 Hz, 2H), 8.27 (s, 1H), 7.81 (d, J = 12.6 Hz, 1H), 7.64 (d, J = 8.5 Hz, 1H), 4.80-4.58 (m, 1H), 4.42 (s, 1H), 4.15-3.80 (m, 2H), 3.41 (d, J = 25.8 Hz, 2H), 3.00 (d, J = 31.7 Hz, 2H), 2.82 (s, 6H), 2.75 (s, 3H), 1.53 (s, 2H), 1.33 (d, J = 20.5 Hz, 3H), 1.19 (s, 2H). LC-MS: m/z(M + H)⁺ = 533.2 I-22 33

Yellow solid, yield 98% ¹H-NMR (400 MHz, DMSO- d₆) δ 8.78 (d, J = 3.3 Hz, 1H), 8.37 (s, 1H), 8.06 (d, J = 8.7 Hz, 1H), 7.89 (d, J = 11.5 Hz, 1H), 7.74 (d, J = 8.7 Hz, 1H), 4.50 (s, 1H), 4.28 (s, 2H), 3.50 (t, J = 6.1 Hz, 2H), 3.11 (t, J = 6.3 Hz, 2H), 2.79 (s, 3H), 2.22 (d, J = 10.4 Hz, 2H), 2.07- 1.95 (m, 2H), 1.86 (d, J = 12.2 Hz, 2H), 1.52 (d, J = 22.4 Hz, 1H), 1.27 (d, J = 12.2 Hz, 2H), 0.96 (d, J = 6.4 Hz, 3H). LC-MS: m/z: (M + H)⁺ = 489.9. Preparation embodi- ment 4 34

Yellow solid, yield 94%. ¹H NMR (400 MHz, DMSO- d₆) δ 9.01 (br, 2H), 8.70 (d, J = 3.8 Hz, 1H), 8.29 (s, 1H), 8.17 (d, J = 8.6 Hz, 1H), 7.78-7.51 (m, 2H), 4.64 (d, J = 32.0 Hz, 2H), 4.39 (s, 1H), 4.17 (d, J = 19.2 Hz, 2H), 3.80 (dt, J = 56.6, 5.8 Hz, 2H), 2.91 (d, J = 44.2 Hz, 2H), 2.66 (s, 3H), 2.58 (d, J = 3.1 Hz, 3H), 2.24 (d, J = 12.0 Hz, 2H), 1.92 (dd, J = 28.7, 12.4 Hz, 3H), 1.52 (s, 1H), 1.34-1.21 (m, 2H), 1.04- 0.90 (m, 3H). LC-MS: m/z: (M + H)⁺ = 560.9. I-4 35

Solid. ¹H-NMR (400 MHz, DMSO- d₆) δ = 10.08-10.10 (m, 1H), 8.70 (d, 1H, J = 4 Hz), 8.29 (s, 1H), 8.13-8.17 (m, 1H), 7.68 (d, 1H, J = 12.4 Hz), 7.54-7.59 (m, 1H), 4.73 (s, 1H), 4.61 (s, 1H), 4.35-4.38 (m, 1H), 3.79- 3.87 (m, 2H), 2.90-2.94 (m, 1H), 2.79-2.82 (m, 1H), 2.66 (s, 4H), 2.22-2.29 (m, 8H), 1.86- 1.97 (m, 4H), 1.21-1.29 (m, 3H), 0.95-0.99 (m, 3H). LC-MS: m/z: (M + H)⁺ = 575.2. I-5 36

Solid. ¹H-NMR(400 MHz, CDCl₃) δ = 8.44 (d, 1H, J = 3.6 Hz), 8.31 (d, 1H, J = 8.4 Hz), 8.20-8.22 (m, 1H), 7.98-8.03 (m, 1H), 7.76-7.81 (m, 1H), 7.46-7.50 (m, 1H), 4.74-4.80 (m, 2H), 4.22-4.29 (m, 1H), 3.92-3.98 (m, 2H), 3.29 (s, 2H), 2.94-3.02 (m, 2H), 2.72 (s, 3H), 2.37 (s, 6H), 2.22-2.33 (m, 2H), 2.02- 2.08 (m, 4H), 1.86-1.92 (m, 1H), 1.53-1.59 (m, 2H). LC-MS: m/z: (M + H)⁺ = 561.2. I-5 38

Solid. ¹H-NMR(400 MHz, DMSO- d₆) δ = 10.12-10.14 (m, 1H), 8.70 (d, 1H, J = 4 Hz), 8.32 (s, 1H), 8.14 (d, 1H, J = 8 Hz), 7.58-7.72 (m, 2H), 6.90-6.98 (m, 1H), 6.18 (dd, 1H, J = 16.8 Hz, 2 Hz), 5.75 (dd, 1H, J = 10.4 Hz, 2.4 Hz), 4.82-4.88 (m, 1H), 4.68-4.79 (m, 2H), 3.87- 3.92 (m, 2H), 2.84-2.92(m, 2H), 2.66(s, 3H), 1.64 (d, 6H, J = 7.2 Hz), 1.24 (s, 3H). LC-MS: m/z: (M + H)⁺ = 490.2. I-10 39

Yellow solid 760 mg, yield 98%. ¹H NMR (400 MHz, DMSO- d₆) δ 9.82 (s, 2H), 8.83 (d, J = 3.4 Hz, 1H), 8.41 (s, 1H), 8.12 (d, J = 8.7 Hz, 1H), 7.92 (d, J = 11.6 Hz, 1H), 7.80 (d, J = 8.7 Hz, 1H), 5.27-5.12 (m, 1H), 4.27 (s, 2H), 3.48 (s, 2H), 3.14 (d, J = 5.9 Hz, 2H), 2.93-2.82 (m, 2H), 2.76 (s, 3H), 2.71- 2.61 (m, 2H), 2.07-1.89 (m, 3H). LC-MS: m/z: (M + H)⁺ = 447.9. I-28 40

Solid. ¹H NMR (400 MHz, DMSO- d₆) δ 10.05 (d, J = 11.9 Hz, 1H), 8.69 (d, J = 3.4 Hz, 1H), 8.31 (s, 1H), 8.13 (d, J = 8.3 Hz, 1H), 7.81-7.51 (m, 2H), 5.00- 4.78 (m, 1H), 4.59 (d, J = 25.3 Hz, 2H), 3.76 (d, J = 39.2 Hz, 2H), 3.65-3.46 (m, 2H), 2.83 (d, J = 25.6 Hz, 2H), 2.65 (s, 3H), 2.46 (d, J = 8.6 Hz, 2H), 2.25 (dd, J = 22.7, 9.9 Hz, 2H), 1.64 (d, J = 6.7 Hz, 6H). LC-MS: m/z: (M + H)⁺ = 532.9. I-4 41

Solid. ¹H NMR (400 MHz, DMSO- d₆) δ 10.90 (s, 1H), 9.88 (br, 1H), 8.93-8.72 (m, 2H), 8.31 (s, 1H), 8.11 (d, J = 8.7 Hz, 1H), 7.83 (d, J = 8.8 Hz, 1H), 5.00 (s, 1H), 4.28 (s, 2H), 3.47 (s, 2H), 3.14 (s, 2H), 2.79 (s, 3H), 1.68 (d, J = 6.9 Hz, 6H). LC-MS: m/z: (M + H)⁺ = 485.9. I-28 42

Yellow solid 5 mg, yield 11%. ¹H-NMR (400 MHz, CD₃OD) δ 8.93 (s, 1H), 8.57 (s, 1H), 8.26-8.15 (m, 2H), 7.55 (s, 1H), 5.14 (s, 2H), 3.99 (d, J = 31.3 Hz, 2H), 3.74 (d, J = 26.0 Hz, 2H), 2.97 (s, 3H), 2.73 (s, 2H), 2.64 (s, 3H), 2.44 (s, 2H), 2.21 (d, J = 7.5 Hz, 1H), 2.04 (s, 1H), 1.83 (d, J = 6.4 Hz, 6H), 1.61 (s, 1H). LC-MS: m/z: (M + H)⁺ = 547.3. I-4 43

Yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 8.55 (d, J = 3.9 Hz, 1H), 8.44 (s, 1H), 8.33-8.20 (m, 2H), 7.84 (d, J = 12.2 Hz, 1H), 7.61 (d, J = 8.8 Hz, 1H), 4.61 (s, 2H), 3.79 (t, J = 5.9 Hz, 2H), 3.55 (t, J = 5.7 Hz, 2H), 3.44- 3.48 (m, 1H), 3.21 (t, J = 5.7 Hz, 2H), 2.95 (t, J = 5.7 Hz, 2H), 2.90 (s, 6H), 2.76 (s, 3H), 1.37 (d, J = 5.3 Hz, 2H), 1.20 (s, 2H). LC-MS: m/z: (M + H)⁺ = 548. I-61 44

Yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 8.55 (d, J = 3.9 Hz, 1H), 8.43 (s, 1H), 8.33 (s, 1H), 8.26 (d, J = 8.5 Hz, 1H), 7.81 (d, J = 11.9 Hz, 1H), 7.58 (d, J = 8.7 Hz, 1H), 5.02-4.88 (m, 1H), 4.60 (s, 2H), 3.79 (t, J = 5.9 Hz, 2H), 3.57 (t, J = 5.6 Hz, 2H), 3.23 (t, J = 6 Hz, 3H), 2.95 (t, J = 5.9 Hz, 3H), 2.92 (s, 6H), 2.71 (s, 3H), 1.74 (d,J = 6.9 Hz, 6H). LC-MS: m/z: (M + H)⁺ = 550. I-61 45

Pale yellow solid. ¹H-NMR(400 MHz, CDCl₃) δ 8.44 (d, 1H, J = 3.6 Hz), 8.19- 8.35 (m, 3H), 7.81(d, 1H, J = 12.0 Hz), 7.46(d, 1H, J = 8.4 Hz), 3.95 (s, 2H), 3.32-3.39 (m, 1H), 3.21-3.31 (m, 1H), 3.12 (s, 4H), 2.76 (s, 3H), 1.35-1.37 (m, 2H), 1.30(d, 6H, J = 6.4 Hz), 1.13-1.17 (m, 2H). LC-MS m/z: (M + H)⁺ = 476.2 I-3 46

Solid. ¹H-NMR(400 MHz, DMSO- d₆) δ = 9.99 (s, 1H), 8.70 (d, 1H, J = 4 Hz), 8.22 (s, 1H), 8.16 (s, 1H), 8.12 (d, 1H, J = 8.4 Hz), 4.56 (s, 2H), 4.15 (t, 2H, J = 11.6 Hz), 3.71-3.75 (m, 2H), 3.41-3.49 (m, 2H), 2.82-2.86 (m, 2H), 2.67 (s, 4H), 2.53-2.56 (m, 3H), 2.22 (s, 6H), 1.24-1.28 (m, 3H), 1.12-1.15 (m, 2H). LC-MS: m/z: (M + H)⁺ = 549.2. I-61 47

Solid. ¹H-NMR(400 MHz, DMSO- d₆) δ = 9.97 (d, 1H, J = 7.2 Hz), 8.69 (d, 1H, J = 4 Hz), 8.21 (s, 1H), 8.10 (d, 1H, J = 8.8 Hz), 7.75 (d, 1H, J = 12 Hz), 7.61- 7.66 (m, 1H), 4.56-4.86 (m, 2H), 3.66-3.94 (m, 3H), 3.44- 3.48 (m, 1H), 2.79-2.91 (m, 2H), 2.67 (s, 3H), 2.17 (d, 1H, J = 14.4 Hz), 1.26-1.30 (m, 2H), 1.13 (s, 2H), 1.05 (d, 3H, J = 6.4 Hz). LC-MS: m/z: (M + H)⁺ = 533.2. I-22 48

Yellow powder. ¹H NMR (400 MHz, CD₃OD) δ 8.97 (s, 1H), 8.57 (s, 1H), 8.25 (t, J = 8.6 Hz, 2H), 7.57 (d, J = 14.1 Hz, 1H), 5.04-4.94 (m, 2H), 4.64 (s, 2H), 4.07 (d, J = 31.7 Hz, 1H), 3.68 (d, J = 47.0 Hz, 2H), 3.04 (s, 3H), 2.73 (d, J = 17.8 Hz, 4H), 1.56 (dd, J = 18.3, 6.7 Hz, 7H). LC-MS: m/z: (M + H)⁺ = 519.2. I-22 49

Solid. ¹H NMR (400 MHz, CD₃OD) δ 8.55 (s, 1H), 8.35 (s, 1H), 8.24 (d, J = 8.2 Hz, 1H), 7.81 (d, J = 11.8 Hz, 1H), 7.58 (d, J = 8.2 Hz, 1H), 5.36 (s, 1H), 3.77 (t, J = 5.6 Hz, 2H), 3.64 (t, J = 5.8 Hz, 2H), 3.18-2.88 (m, 4H), 2.71 (s, 3H), 2.14 (d, J = 62.1 Hz, 4H), 1.74 (d, J = 6.9 Hz, 6H). LC-MS: m/z: (M + H)⁺ = 522.9. I-61 50

Solid. ¹H NMR (400 MHz, DMSO- d₆) δ 9.96 (s, 1H), 8.69 (d, J = 3.9 Hz, 1H), 8.22 (s, 1H), 8.11 (d, J = 8.4 Hz, 1H), 7.75 (d, J = 11.7 Hz, 1H), 7.57 (d, J = 8.5 Hz, 1H), 6.63 (s, 1H), 5.33 (s, 1H), 4.56 (d, J = 54.3 Hz, 4H), 3.67 (t, J = 5.7 Hz, 2H), 3.14 (d, J = 5.8 Hz, 2H), 2.81 (s, 2H), 2.67 (s, 3H), 2.14-1.79 (m, 4H). LC-MS: m/z: (M + H)⁺ = 520.9. I-61 51

Solid. ¹H NMR (400 MHz, DMSO- d₆) δ 9.07 (s, 2H), 8.88 (t, J = 3.2 Hz, 1H), 8.38 (s, 1H), 8.09 (dd, J = 8.8, 3.4 Hz, 1H), 7.94 (dd, J = 45.3, 10.2 Hz, 2H), 5.22-5.13 (m, 1H), 4.69 (d, J = 25.5 Hz, 2H), 4.20 (dd, J = 17.8, 12.4 Hz, 2H), 3.83 (dt, J = 52.4, 5.7 Hz, 2H), 3.04 (d, J = 51.3 Hz, 2H), 2.82 (dd, J = 20.4, 10.6 Hz, 2H), 2.73 (s, 3H), 2.65 (dd, J = 16.9, 7.9 Hz, 2H), 2.58 (d, J = 5.3 Hz, 3H), 1.99 (dd, J = 23.4, 10.6 Hz, 2H). LC-MS: m/z: (M + H)⁺ = 518.9. I-4 53

Solid. ¹H NMR (400 MHz, DMSO- d₆) δ 10.00 (d, J = 10.1 Hz, 1H), 8.70 (s, 1H), 8.18 (d, J = 29.5 Hz, 2H), 7.87-7.57 (m, 2H), 4.67 (d, J = 23.5 Hz, 2H), 3.82 (s, 2H), 3.47 (s, 2H), 3.33 (s, 5H), 2.88 (d, J = 49.8 Hz, 6H), 2.67 (s, 3H), 1.27 (s, 2H), 1.13 (s, 2H). LC-MS: m/z: (M + H)⁺ = 532.9. I-6 54

Solid. ¹H-NMR(400 MHz, DMSO- d₆) δ = 10.02-10.04 (m, 1H), 8.70 (d, 1H, J = 3.6 Hz), 8.35 (s, 1H), 8.11-8.15 (m, 1H), 7.73 (d, 1H, J = 12 Hz), 7.58-7.61 (m, 1H), 5.32-5.35 (m, 1H), 5.07-5.12 (m, 1H), 4.61-4.72 (m, 2H), 3.80-3.86 (m, 2H), 2.81-2.93 (m, 4H), 2.30 (s, 6H), 1.95-2.04 (m, 6H), 1.44-1.48 (m, 2H), 0.84-0.88 (m, 2H). LC-MS: m/z: (M + H)⁺ = 533.2. I-5 55

Solid. ¹H NMR (400 MHz, DMSO- d₆) δ 10.06 (d, J = 16.5 Hz, 1H), 8.70 (d, J = 3.8 Hz, 1H), 8.31 (s, 1H), 8.19-8.05 (m, 1H), 7.86-7.52 (m, 2H), 4.96- 4.76 (m, 1H), 4.62 (dd, J = 20.8, 13.4 Hz, 2H), 3.77 (d, J = 33.9 Hz, 2H), 3.20 (d, J = 9.0 Hz, 2H), 2.84 (d, J = 32.0 Hz, 2H), 2.65 (s, 3H), 2.56 (s, 6H), 2.46-2.18 (m, 4H), 1.64 (d, J = 6.8 Hz, 6H). LC-MS: m/z: (M + H)⁺ = 560.9. I-22 57

Yellow powder. ¹H NMR (400 MHz, CD₃OD) δ 8.97 (d, J = 3.0 Hz, 1H), 8.58 (s, 1H), 8.25 (dd, J = 21.3, 10.1 Hz, 2H), 7.63 (d, J = 9.1 Hz, 1H), 4.59-4.46 (m, 4H), 3.75 (t, J = 6.1 Hz, 2H), 3.53 (t, J = 6.0 Hz, 2H), 3.03 (s, 3H), 1.51- 1.42 (m, 1H), 0.83-0.73 (m, 2H), 0.67 (q, J = 5.1 Hz, 2H). LC-MS: m/z: (M + H)⁺ = 448.2. I-28 58

Yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 8.53 (s, 1H), 8.42 (d, J = 16.9 Hz, 1H), 7.93 (d, J = 30.5 Hz, 1H), 7.72 (d, J = 27.6 Hz, 1H), 7.51-7.08 (m, 2H), 4.62 (s, 2H), 3.90 (d, J = 32.8 Hz, 1H), 3.77 (d, J = 39.8 Hz, 2H), 3.17 (d, J = 22.1 Hz, 2H), 2.91 (d, J = 12.1 Hz, 3H), 2.73 (s, 2H), 2.55 (d, J = 36.5 Hz, 3H), 2.16 (d, J = 72.2 Hz, 4H), 1.19 (s, 2H), 1.05-0.69 (m, 2H). LC-MS: m/z: (M + H)⁺ = 544.9. I-22 59

Solid. ¹H-NMR(400 MHz, DMSO- d₆) δ9.98-10.00 (m, 1H), 8.69 (d, 1H, J = 4 Hz), 8.20-8.21 (m, 2H), 8.12 (d, 1H, J = 8.8 Hz), 7.75 (d, 1H, J = 12.4 Hz), 7.62- 7.67 (m, 1H), 4.55-4.82 (m, 2H), 3.80-4.04 (m, 2H), 3.33- 3.48 (m, 3H), 3.02-3.06 (m, 1H), 2.80-2.90 (m, 2H), 2.67 (s, 3H), 2.28-2.33 (m, 4H), 1.79 (s, 2H), 1.24-1.30 (m, 3H), 1.12- 1.14 (m, 2H). LC-MS: m/z: (M + H)⁺ = 545.2. I-22 60

Yellow powder. ¹H NMR (400 MHz, CD₃OD) δ 8.49 (d, J = 33.2 Hz, 1H), 8.19 (d, J = 29.6 Hz, 1H), 7.79 (s, 1H), 7.60 (s, 1H), 4.92 (d, J = 13.2 Hz, 1H), 4.70 (d, J = 33.6 Hz, 2H), 4.18 (d, J = 41.1 Hz, 4H), 3.88 (d, J = 63.1 Hz, 2H), 2.98 (d, J = 36.9 Hz, 2H), 2.85 (s, 6H), 2.71 (s, 3H), 0.74- 0.42 (m, 4H). LC-MS: m/z: (M + H)⁺ = 533.2. I-5 63

Solid. ¹H-NMR(DMSO-d₆) δ 9.92 (s, 1H), 8.68 (d, 1H, J = 4 Hz), 8.31 (d, 1H, J = 1.2 Hz), 8.06 (d, 1H, J = 8.4 Hz), 7.70 (d, 1H, J = 12 Hz), 7.45 (d, 1H, J = 8.4 Hz), 4.82-4.87 (m, 1H), 3.58 (s, 2H), 2.80-2.84 (m, 4H), 2.65 (s, 3H), 2.58-2.63 (m, 3H), 2.22 (s, 6H), 1.64 (d, 6H, J = 6.8 Hz). LC-MS: m/z: (M + H)⁺ = 507.9. I-52 64

Pale yellow solid. ¹H-NMR(400 MHz, CD₃OD) δ 8.44-8.45 (m, 1H), 8.35-8.39 (m, 1H), 7.93 (s, 1H), 7.59- 7.62(m, 1H), 7.52 (s, 1H), 4.65- 4.74 (m, 2H), 3.80-3.98 (m, 4H), 3.37-3.38 (m, 1H), 2.94- 3.03 (m, 2H), 2.75 (s, 6H), 2.71 (s, 3H), 1.30-1.34 (m, 2H), 1.15-1.17 (m, 2H). LC-MS: m/z: (M + H)⁺ = 517.2. I-5 65

Solid. ¹H NMR (400 MHz, CD₃OD) δ 8.97 (t, J = 3.2 Hz, 1H), 8.58 (s, 1H), 8.25 (dd, J = 12.0, 9.0 Hz, 2H), 7.57 (dd, J = 8.7, 4.8 Hz, 1H), 4.97 (dd, J = 17.2, 6.6 Hz, 2H), 4.81 (d, J = 17.6 Hz, 1H), 4.18 (dd, J = 108.9, 17.0 Hz, 2H), 3.86 (dd, J = 30.6, 26.8 Hz, 2H), 3.57-3.40 (m, 2H), 3.28 (s, 1H), 3.05 (s, 3H), 2.76- 1.90 (m, 4H), 1.67-1.48 (m, 2H), 1.35 (d, J = 30.5 Hz, 2H). LC-MS: m/z: (M + H)⁺ = 530.9. I-4 66

White powder. ¹H NMR (400 MHz, CD₃OD) δ 8.97 (s, 1H), 8.57 (s, 1H), 8.25 (t, J = 8.6 Hz, 2H), 7.57 (d, J = 14.1 Hz, 1H), 5.04-4.94 (m, 2H), 4.64 (s, 2H), 4.07 (d, J = 31.7 Hz, 1H), 3.68 (d, J = 47.0 Hz, 2H), 3.04 (s, 3H), 2.73 (d, J = 17.8 Hz, 4H), 1.56 (dd, J = 18.3, 6.7 Hz, 7H). LC-MS: m/z: (M + H)⁺ = 519.2. I-4 67

Solid. ¹H NMR (400 MHz, CD₃OD) δ 8.54 (dd, J = 9.3, 7.7 Hz, 2H), 8.29 (d, J = 7.9 Hz, 1H), 7.82 (d, J = 12.3 Hz, 1H), 7.65 (dd, J = 8.7, 4.6 Hz, 1H), 4.74 (dd, J = 33.1, 18.6 Hz, 4H), 4.00- 3.82 (m, 2H), 3.51-3.43 (m, 2H), 3.04 (d, J = 6.2 Hz, 1H), 2.96 (s, 1H), 2.75 (s, 3H), 2.16- 1.91 (m, 4H), 1.41-1.30 (m, 4H). LC-MS: m/z: (M + H)⁺ = 530.9. I-4 68

Pale yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 8.96 (d, J = 2.9 Hz, 1H), 8.76 (s, 1H), 8.22 (dd, J = 9.6, 7.2 Hz, 2H), 7.65 (d, J = 8.9 Hz, 1H), 4.52 (s, 2H), 3.75 (s, 2H), 3.53 (s, 2H), 3.16 (s, 3H), 2.07 (s, 9H). LC-MS: m/z: (M + H)⁺ = 449.9. I-28 69

Pale yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 8.45-8.39 (m, 2H), 8.24 (d, J = 8.4 Hz, 1H), 7.94 (s, 1H), 7.77 (d, J = 11.5 Hz, 1H), 7.37 (d, J = 8.5 Hz, 1H), 3.70 (s, 2H), 2.99 (t, J = 5.8 Hz, 2H), 2.94- 2.88 (m, 5H), 2.82 (d, J = 6.0 Hz, 2H), 2.77-2.69 (m, 2H), 2.46 (s, 6H), 1.94 (s, 9H). LC-MS: m/z: (M + H)⁺ = 521.0. I-52 70

Yellow powder. ¹H NMR (400 MHz, CDCl₃) δ 8.40 (d, J = 3.7 Hz, 1H), 8.23- 8.15 (m, 2H), 7.95 (s, 1H), 7.79 (d, J = 11.7 Hz, 1H), 7.39 (d, J = 8.5 Hz, 1H), 3.62 (s, 2H), 3.35-3.26 (m, 1H), 2.95 (d, J = 5.8 Hz, 2H), 2.84 (t, J = 5.9 Hz, 2H), 2.73 (s, 3H), 2.62 (dd, J = 12.8, 6.0 Hz, 4H), 2.43 (s, 6H), 1.91 (dd, J = 14.9, 7.4 Hz, 2H), 1.36-1.31 (m, 2H), 1.14 (d, J = 7.2 Hz, 2H). LC-MS: m/z: (M + H)⁺ = 519.3. I-3 71

Solid. ¹H NMR (400 MHz, CDCl₃) δ 8.43 (d, J = 3.8 Hz, 1H), 8.23 (t, J = 5.1 Hz, 2H), 7.99 (s, 1H), 7.80 (d, J = 11.6 Hz, 1H), 7.38 (d, J = 8.5 Hz, 1H), 4.75 (dt, J = 14.0, 7.0 Hz, 1H), 3.75 (d, J = 15.3 Hz, 2H), 3.53-3.46 (m, 2H), 3.00 (d, J = 5.2 Hz, 2H), 2.96 (d, J = 5.1 Hz, 2H), 2.77 (s, 4H), 2.71 (s, 3H), 2.42 (s, 3H), 2.28 (t, J = 2.3 Hz, 1H), 1.73 (d, J = 7.0 Hz, 6H). LC-MS: m/z: (M + H)⁺ = 530.9. I-52 74

Yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 8.53 (d, J = 3.9 Hz, 1H), 8.27 (s, 1H), 8.18 (d, J = 8.6 Hz, 1H), 7.81 (d, J = 12.0 Hz, 1H), 7.54 (d, J = 8.5 Hz, 1H), 3.76 (dd, J = 35.1, 14.8 Hz, 4H), 3.62-3.56 (m, 1H), 3.46 (m, 1H), 3.15-3.06 (m, 2H), 3.02- 2.93 (m, 5H), 2.93-2.85 (m, 2H), 2.81-2.73 (m, 3H), 2.35 (m, 1H), 2.22-2.00 (m, 2H), 1.88-1.75 (m, 1H), 1.37 (m, 2H), 1.22-1.16 (m, 2H). LC-MS m/z: (M + H)⁺ = 530.9. I-3 75

Yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 8.39 (d, J = 3.8 Hz, 1H), 8.21 (d, J = 8.5 Hz, 2H), 7.89 (s, 1H), 7.78 (d, J = 12.0 Hz, 1H), 7.36 (d, J = 8.4 Hz, 1H), 4.71 (m, 1H), 3.65 (s, 4H), 2.93 (d, J = 3.7 Hz, 2H), 2.86 (d, J = 11.6 Hz, 2H), 2.69 (s, 3H), 2.61 (s, 2H), 2.21 (d, J = 12.0 Hz, 1H), 2.00 (s, 1H), 1.71 (d, J = 6.9 Hz, 6H), 1.30 (d, J = 11.4 Hz, 6H). LC-MS: m/z: (M + H)⁺ = 557.3. I-72 76

Yellow powder. ¹H NMR (400 MHz, DMSO- d₆) δ 9.93 (s, 1H), 8.68 (d, J = 3.9 Hz, 1H), 8.31 (s, 1H), 8.07 (d, J = 8.5 Hz, 1H), 7.70 (d, J = 12.2 Hz, 1H), 7.50 (d, J = 8.5 Hz, 1H), 4.96-4.78 (m, 1H), 3.65 (d, J = 3.5 Hz, 2H), 3.51 (d, J = 2.0 Hz, 2H), 3.23 (s, 1H), 2.85 (dd, J = 10.4, 4.3 Hz, 4H), 2.65 (s, 3H), 1.64 (d, J = 6.9 Hz, 6H). LC-MS: m/z: (M + H)⁺ = 473.9. I-3 78

Yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 8.97 (s, 1H), 8.58 (s, 1H), 8.28 (s, 1H), 8.15 (s, 1H), 7.51 (s, 1H), 4.10 (s, 2H), 3.64 (dt, J = 9.4, 4.4 Hz, 4H), 3.37 (s, 2H), 3.02 (d, J = 24.1 Hz, 4H), 1.50 (d, J = 35.2 Hz, 4H), 1.42- 1.27 (m, 6H). LC-MS: m/z: (M + H)⁺ = 504.9. I-77 79

Yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 8.52 (d, J = 3.9 Hz, 1H), 8.29 (s, 1H), 8.15 (d, J = 8.5 Hz, 1H), 7.72 (d, J = 11.7 Hz, 1H), 7.45 (d, J = 8.4 Hz, 1H), 4.91 (d, J = 7.1 Hz, 1H), 3.75 (s, 2H), 3.60 (s, 2H), 3.40 (s, 2H), 2.97 (s, 4H), 2.69 (s, 3H), 2.38 (s, 6H), 1.72 (d, J = 6.9 Hz, 6H). LC-MS: m/z: (M + H)⁺ = 530.9. I-3 81

Yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 8.95 (d, J = 3.1 Hz, 1H), 8.55 (s, 1H), 8.23 (t, J = 8.1 Hz, 1H), 8.18-8.13 (m, 1H), 7.55 (d, J = 8.9 Hz, 1H), 4.29 (s, 2H), 3.90 (s, 1H), 3.72 (s, 2H), 3.51 (s, 4H), 3.02 (s, 3H), 1.52 (d, J = 7.2 Hz, 2H), 1.45 (d, J = 6.3 Hz, 2H), 1.35 (s, 3H), 1.29 (s, 1H). LC-MS: m/z: (M + H)⁺ = 491.2. I-52 82

Solid. ¹H NMR (400 MHz, DMSO- d₆) δ 9.95 (s, 1H), 8.67 (d, J = 3.8 Hz, 1H), 8.26 (s, 1H), 8.05 (d, J = 8.5 Hz, 1H), 7.71 (d, J = 12.0 Hz, 1H), 7.43 (d, J = 8.4 Hz, 1H), 4.35 (s, 1H), 3.88 (s, 2H), 3.09 (t, J = 5.6 Hz, 2H), 2.76 (t, J = 5.8 Hz, 2H), 2.64 (s, 4H), 2.20-2.07 (m, 2H), 2.06- 1.93 (m, 2H), 0.74 (t, J = 7.3 Hz, 7H). LC-MS: m/z: (M + H)⁺ = 464.9. I-39 83

Pale yellow solid. ¹H NMR (400 MHz, DMSO- d₆) δ 9.95 (s, 1H), 8.67 (d, J = 3.9 Hz, 1H), 8.26 (s, 1H), 8.05 (d, J = 8.4 Hz, 1H), 7.70 (d, J = 12.4 Hz, 1H), 7.44 (d, J = 8.5 Hz, 1H), 4.39-4.28 (m, 1H), 3.58 (s, 2H), 2.82 (dd, J = 9.4, 4.2 Hz, 5H), 2.66-2.57 (m, 7H), 2.22 (s, 7H), 2.13 (dd, J = 16.9, 7.2 Hz, 3H), 2.06-1.95 (m, 3H), 0.74 (t, J = 7.3 Hz, 7H). LC-MS: m/z: (M + H)⁺ = 535.0. I-52 84

White solid. ¹H NMR (400 MHz, CD₃OD) δ 8.88 (d, J = 3.3 Hz, 1H), 8.66 (s, 1H), 8.20 (d, J = 11.1 Hz, 1H), 8.07 (d, J = 8.9 Hz, 1H), 7.79 (d, J = 8.9 Hz, 1H), 4.48 (s, 2H), 4.38-4.25 (m, 1H), 3.71 (dd, J = 10.9, 4.8 Hz, 2H), 3.44 (d, J = 6.0 Hz, 2H), 2.95 (s, 3H), 1.84 (t, J = 7.0 Hz, 3H), 1.78 (dd, J = 11.8, 5.4 Hz, 1H), 0.93 (m, 1H), 0.82-0.74 (m, 1H), 0.69-0.60 (m, 1H), 0.50- 0.38 (m, 1H). LC-MS: m/z: (M + H)⁺ = 462.0. I-28 85

Yellow powder. ¹H NMR (400 MHz, CDCl₃) δ 8.53 (s, 1H), 8.43 (s, 1H), 8.24 (d, J = 8.5 Hz, 1H), 8.19 (s, 1H), 7.80 (d, J = 11.6 Hz, 1H), 7.41 (d, J = 8.5 Hz, 1H), 3.70 (s, 2H), 3.40 (dt, J = 10.5, 8.3 Hz, 2H), 2.99-2.95 (m, 4H), 2.76 (s, 3H), 2.69 (s, 6H), 2.60- 2.50 (m, 1H), 1.40-1.35 (m, 2H), 1.32 (s, 3H), 1.27 (s, 1H), 1.15 (s, 2H). LC-MS: m/z: (M + H)⁺ = 519.3. I-80 86

Pale yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 8.54 (d, J = 3.9 Hz, 1H), 8.40 (d, J = 1.1 Hz, 1H), 8.18 (d, J = 8.5 Hz, 1H), 7.81 (d, J = 11.9 Hz, 1H), 7.50 (d, J = 8.5 Hz, 1H), 4.00 (dq, J = 14.0, 7.0 Hz, 1H), 3.74 (s, 2H), 3.03-2.95 (m, 6H), 2.85 (t, J = 6.6 Hz, 2H), 2.67 (s, 3H), 2.62 (s, 6H), 1.77 (d, J = 7.0 Hz, 3H), 1.75-1.67 (m, 1H), 0.86 (m, 1H), 0.65 (m, 1H), 0.61-0.53 (m, 1H), 0.37- 0.28 (m, 1H). LC-MS: m/z: (M + H)⁺ = 533.3. I-52 87

yellow solid. ¹H NMR (400 MHz, DMSO- d₆) δ 9.88 (s, 1H), 8.68 (d, J = 3.9 Hz, 1H), 8.23 (s, 1H), 8.06 (d, J = 8.4 Hz, 1H), 7.73 (d, J = 12.1 Hz, 1H), 7.44 (d, J = 8.4 Hz, 1H), 3.83 (s, 2H), 3.04 (t, J = 5.8 Hz, 2H), 2.73 (t, J = 5.6 Hz, 2H), 2.69 (s, 3H), 1.56 (s, 3H), 1.38-1.13 (m, 4H). LC-MS: m/z: (M + H)⁺ = 447.9. I-28 88

Pale yellow solid. ¹H NMR (400 MHz, DMSO- d₆) δ 9.93 (s, 1H), 8.69 (d, J = 3.9 Hz, 1H), 8.23 (s, 1H), 8.08 (d, J = 8.4 Hz, 1H), 7.74 (d, J = 12.1 Hz, 1H), 7.48 (d, J = 8.5 Hz, 1H), 3.59 (s, 2H), 2.92- 2.77 (m, 4H), 2.69 (s, 3H), 2.61 (dd, J = 15.2, 5.8 Hz, 4H), 2.28 (s, 6H), 1.56 (s, 3H), 1.23 (d, J = 11.8 Hz, 4H). LC-MS: m/z: (M + H)⁺ = 518.9. I-52 89

Yellow solid. ¹H NMR (400 MHz, CD₃OD + CDCl₃) δ 8.52 (d, J = 3.8 Hz, 1H), 8.31 (d, J = 8.6 Hz, 1H), 8.28 (d, J = 1.1 Hz, 1H), 7.77 (d, J = 11.2 Hz, 1H), 7.58 (d, J = 8.7 Hz, 1H), 4.89 (d, J = 6.9 Hz, 1H), 4.25 (s, 2H), 3.49 (t, J = 6.0 Hz, 2H), 3.20 (dd, J = 10.9, 5.4 Hz, 4H), 2.70 (s, 3H), 1.73 (d, J = 6.9 Hz, 6H), 1.18-1.04 (m, 2H), 0.14 (d, J = 3.3 Hz, 9H). LC-MS: m/z: (M + H)⁺ = 536.0. I-3 90

Tawney solid. ¹H NMR (400 MHz, CD₃OD) δ 8.57 (d, J = 3.9 Hz, 1H), 8.51 (s, 1H), 8.21 (d, J = 8.5 Hz, 1H), 7.79 (d, J = 11.6 Hz, 1H), 7.61 (d, J = 8.4 Hz, 1H), 4.03 (s, 2H), 3.25-3.29 (m, 4H), 3.10-3.12 (m, 2H), 3.04-2.97 (m, 2H), 2.95 (s, 6H), 2.89 (s, 3H), 2.23-2.11 (m, 2H), 1.96 (s, 9H). LC-MS m/z: (M + H)⁺ = 534.9. I-3 91

Yellow solid. ¹H NMR (400 MHz, CD₃OD + CDCl₃) δ 8.52 (s, 1H), 8.50 (d, J = 3.9 Hz, 1H), 8.28 (d, J = 1.0 Hz, 1H), 8.18 (d, J = 8.5 Hz, 1H), 7.76 (d, J = 11.9 Hz, 1H), 7.47 (d, J = 8.6 Hz, 1H), 4.90 (d, J = 6.9 Hz, 1H), 3.74 (s, 2H), 3.60 (t, J = 5.2 Hz, 2H), 3.10 (s, 3H), 2.97 (s, 4H), 2.85 (t, J = 5.3 Hz, 2H), 2.70 (s, 3H), 1.73 (d, J = 6.9 Hz, 6H). LC-MS: m/z: (M + H)⁺ = 535.0. I-72 92

Pale yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 8.56-8.52 (m, 1H), 8.51 (s, 1H), 8.25 (dd, J = 12.0, 8.6 Hz, 1H), 7.75 (d, J = 11.7 Hz, 1H), 7.63-7.54 (m, 1H), 4.72 (d, 10.2 Hz, 2H), 3.95-3.86 (m, 2H), 2.98 m, 3H), 2.91-2.80 (m, 6H), 2.52 (d, J = 7.3 Hz, 6H), 1.96 (d, J = 3.3 Hz, 9H). LC-MS m/z: (M + H)⁺ = 548.9. 93

Pale yellow solid. 1H NMR (400 MHz, CD₃OD) δ 8.54 (m, 2H), 8.21 (d, J = 8.4 Hz, 1H), 7.79 (d, J = 11.6 Hz, 1H), 7.51 (d, J = 8.6 Hz, 1H), 3.98-3.83 (m, 2H), 3.77 (d, J = 1.5 Hz, 2H), 3.70 (td, J = 11.7, 2.2 Hz, 2H), 3.04-2.98 (m, 5H), 2.2-2.89 (m, 4H), 2.84-2.71 (m, 2H), 2.66 (dd, J = 13.3, 4.1 Hz, 1H), 2.38 (s, 3H), 2.25 (td, J = 11.8, 3.3 Hz, 1H), 1.97 (s, 9H). LC-MS m/z: (M + H)⁺ = 563.3. I-22 94

Pale yellow solid. 1H NMR (400 MHz, CD₃OD) δ 8.53 (d, J = 4.0 Hz, 1H), 8.51 (d, J = 1.1 Hz, 1H), 8.20 (d, J = 8.5 Hz, 1H), 7.76 (d, J = 11.7 Hz, 1H), 7.50 (d, J = 8.6 Hz, 1H), 4.01 (dd, J = 12.5, 2.7 Hz, 1H), 3.96-3.90 (m, 1H), 3.86 (s, 2H), 3.75 (td, J = 11.9, 2.2 Hz, 1H), 3.10-3.07 (m, 3H), 3.04- 3.0 (m, 3H), 2.95 (d, J = 12.7 Hz, 1H), 2.89 (s, 3H), 2.87- 2.82 (m, 1H), 2.79-2.76 (m, 1H), 2.54-2.35 (m, 4H), 1.96 (s, 9H). LC-MS m/z: (M + H)⁺ = 563.3. I-22 96

Pale yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 8.47 (d, J = 3.8 Hz, 1H), 8.27 (d, J = 1.4 Hz, 1H), 8.21 (d, J = 8.5 Hz, 1H), 7.81-7.74 (m, 1H), 7.45 (d, J = 8.6 Hz, 1H), 4.85 (p, J = 6.9 Hz, 1H), 3.69 (s, 2H), 3.42 (t, J = 7.2 Hz, 2H), 2.98-2.89 (m, 4H), 2.83 (s, 3H), 2.73-2.68 (m, 2H), 2.68 (s, 3H), 2.04 (s, 3H), 1.73 (s, 3H), 1.71 (s, 3H). LC-MS m/z: (M + H)⁺ = 550.3. I-95 98

Pale yellow solid. 1H NMR (400 MHz, CD₃OD) δ 8.52 (d, J = 3.9 Hz, 1H), 8.19 (s, 1H), 8.16 (d, J = 8.4 Hz, 1H), 7.78 (d, J = 12.0 Hz, 1H), 7.48 (d, J = 8.5 Hz, 1H), 5.05 (dd, J = 17.6, 8.8 Hz, 1H), 3.70 (s, 2H), 2.95 (dd, J = 10.0, 4.4 Hz, 4H), 2.78 (s, 4H), 2.70 (s, 3H), 2.44 (s, 6H), 2.31-2.22 (m, 4H), 2.08 (td, J = 12.9, 6.7 Hz, 2H), 1.90 (dt, J = 9.6, 3.4 Hz, 2H). LC-MS m/z: (M + H)⁺ = 533.3. I-52 101

White solid. 1H NMR (400 MHz, CD₃OD) δ 8.98 (d, J = 3.2 Hz, 1H), 8.76 (s, 1H), 8.34 (d, J = 9.2 Hz, 1H), 8.24 (d, J = 10.8 Hz, 1H), 7.65 (d, J = 9.1 Hz, 1H), 4.84- 4.79 (m, 1H), 3.83 (dt, J = 11.0, 5.7 Hz, 1H), 3.74-3.66 (m, 1H), 3.62-3.49 (m, 2H), 3.17 (s, 3H), 2.07 (s, 9H), 1.81 (d, J = 6.8 Hz, 3H). LC-MS m/z: (M + H)⁺ = 464.3. I-28

102

Yellow solid. 1H NMR (400 MHz, CD₃OD) δ 8.52 (d, J = 4.0 Hz, 1H), 8.32 (s, 1H), 8.15 (d, J = 8.4 Hz, 1H), 7.72 (d, J = 12.1 Hz, 1H), 7.44 (d, J = 8.2 Hz, 1H), 4.42 (t, J = 12.6 Hz, 1H), 3.66 (s, 2H), 2.92 (dd, J = 13.7, 4.3 Hz, 4H), 2.78-2.68 (m, 7H), 2.40 (s, 6H), 2.31-2.22 (m, 2H), 2.02 (d, J = 11.2 Hz, 4H), 1.84 (d, J = 14.7 Hz, 1H), 1.67-1.57 (m, 2H), 1.42-1.27 (m, 2H). LC-MS m/z: (M + H)⁺ = 547.3. I-52 103

White solid. 1H NMR (400 MHz, CD₃OD) δ 8.54 (d, J = 3.9 Hz, 1H), 8.51 (s, 1H), 8.39 (s, 1H), 8.20 (d, J = 8.6 Hz, 1H), 7.78 (d, J = 11.6 Hz, 1H), 7.57 (d, J = 8.6 Hz, 1H), 4.04 (q, J = 6.6 Hz, 1H), 3.39-3.35 (m, 1H), 3.30-3.23 (m, 1H), 3.12-2.95 (m, 5H), 2.93 (s, 6H), 2.89 (s, 3H), 2.84- 2.76 (m, 1H), 1.96 (s, 9H), 1.43 (d, J = 6.7 Hz, 3H). LC-MS m/z: (M + H)⁺ = 535.3. I-52

104

Yellow solid. 1H NMR (400 MHz, CD₃OD) δ 8.98 (d, J = 3.2 Hz, 1H), 8.76 (s, 1H), 8.34 (d, J = 9.2 Hz, 1H), 8.24 (d, J = 10.8 Hz, 1H), 7.65 (d, J = 9.1 Hz, 1H), 4.84- 4.79 (m, 1H), 3.83 (dt, J = 11.0, 5.7 Hz, 1H), 3.74-3.66 (m, 1H), 3.62-3.49 (m, 2H), 3.17 (s, 3H), 2.07 (s, 9H), 1.81 (d, J = 6.8 Hz, 3H). LC-MS m/z: (M + H)⁺ = 464.3. I-28

106

Yellow solid. 1H NMR (400 MHz, CD₃OD) δ 8.53 (d, J = 4.0 Hz, 1H), 8.51 (d, J = 1.1 Hz, 1H), 8.20 (d, J = 8.5 Hz, 1H), 7.78 (d, J = 11.5 Hz, 1H), 7.57 (d, J = 8.6 Hz, 1H), 4.02 (q, J = 6.6 Hz, 1H), 3.29-3.22 (m, 1H), 3.17 (dd, J = 9.6, 5.9 Hz, 2H), 3.00 (m, 4H), 2.89 (s, 3H), 2.85-2.81 (m, 1H), 2.78 (s, 6H), 1.96 (s, 9H), 1.42 (d, J = 6.7 Hz, 3H). LC-MS m/z: (M + H)⁺ = 535.3. I-52

108

Yellow solid. 1H NMR (400 MHz, CD₃OD) δ 8.54 (d, J = 3.8 Hz, 1H), 8.36 (s, 1H), 8.21 (d, J = 8.4 Hz, 1H), 7.78 (d, J = 11.6 Hz, 1H), 7.50 (d, J = 8.2 Hz, 1H), 4.43 (t, J = 13.2 Hz, 1H), 3.74 (s, 2H), 3.03-2.95 (m, 6H), 2.86 (t, J = 6.5 Hz, 2H), 2.71 (s, 3H), 2.63 (s, 6H), 2.38 (dd, J = 24.7, 10.6 Hz, 2H), 2.09-1.94 (m, 4H), 1.67-1.50 (m, 1H), 1.44- 1.27 (m, 2H), 1.05 (d, J = 6.4 Hz, 3H). LC-MS m/z: (M + H)⁺ = 561.3. I-52 109

White solid. 1H NMR (400 MHz, CD₃OD) δ 8.96 (s, 1H), 8.76 (s, 1H), 8.23 (d, J = 10.1 Hz, 2H), 7.63 (d, J = 8.5 Hz, 1H), 4.94 (s, 1H), 4.64-4.48 (m, 2H), 3.93 (s, 1H), 3.62-3.59 (m, 1H), 3.16 (s, 3H), 2.07 (s, 9H), 1.63 (d, J = 2.1 Hz, 3H). LC-MS m/z: (M + H)⁺ = 464.3. I-28

111

Yellow solid. 1H NMR (400 MHz, CD₃OD) δ 8.97 (d, J = 3.2 Hz, 1H), 8.77 (s, 1H), 8.26 (s, 1H), 8.23 (d, J = 2.9 Hz, 1H), 7.62 (d, J = 9.0 Hz, 1H), 4.63-4.50 (m, 2H), 3.99-3.88 (m, 1H), 3.64-3.60 (m, 1H), 3.41-3.35 (m, 1H), 3.17 (s, 3H), 2.07 (s, 9H), 1.64 (d, J = 6.4 Hz, 3H). LC-MS m/z: (M + H)⁺ = 464.2. I-28

112

White solid. 1H NMR (400 MHz, CD₃OD) δ 8.55 (d, J = 3.9 Hz, 1H), 8.52 (d, J = 1.2 Hz, 1H), 8.47 (s, 1H), 8.22 (d, J = 8.4 Hz, 1H), 7.80 (d, J = 12.0 Hz, 1H), 7.54 (d, J = 8.5 Hz, 1H), 3.88 (s, 2H), 3.41-3.37 (m, 2H), 3.28 (dd, J = 8.2, 4.8 Hz, 1H), 3.01- 2.97 (m, 2H), 2.91 (m, 10H), 2.69 (dd, J = 17.2, 4.9 Hz, 1H), 1.97 (s, 9H), 1.21 (d, J = 6.6 Hz, 3H). LC-MS m/z: (M + H)⁺ = 535.3. I-52

114

Yellow solid. 1H NMR (400 MHz, CD₃OD) δ 8.55 (d, J = 4.0 Hz, 1H), 8.52 (d, J = 1.1 Hz, 1H), 8.20 (d, J = 8.5 Hz, 1H), 7.79 (d, J = 11.6 Hz, 1H), 7.56 (d, J = 8.5 Hz, 1H), 3.89 (s, 2H), 3.42-3.35 (m, 3H), 3.17 (dd, J = 17.2, 5.3 Hz, 1H), 3.00 (td, J = 5.2, 1.4 Hz, 2H), 2.93 (s, 6H), 2.90 (s, 3H), 2.69 (dd, J = 17.2, 4.7 Hz, 1H), 1.97 (s, 9H), 1.21 (d, J = 6.6 Hz, 3H). LC-MS m/z: (M + H)⁺ = 535.3. I-52

126

Yellow powder. ¹H NMR (400 MHz, CD₃OD) δ 8.52 (s, 1H), 8.15 (d, J = 7.7 Hz, 2H), 7.80 (d, J = 12.3 Hz, 1H), 7.52 (d, J = 6.0 Hz, 1H), 4.23 (d, J = 6.8 Hz, 2H), 3.70 (s, 2H), 2.94 (d, J = 9.3 Hz, 4H), 2.79 (s, 3H), 2.69 (s, 4H), 2.47 (s, 6H), 0.89 (t, J = 7.4 Hz, 1H), 0.66 (d, J = 3.0 Hz, 2H), 0.51 (d, J = 10.7 Hz, 2H). LC-MS: m/z: (M + H)⁺ = 519.3. I-52 127

Yellow powder. ¹H NMR (400 MHz, CD₃OD) δ 8.47 (d, J = 51.2 Hz, 2H), 8.27 (s, 1H), 7.81 (d, J = 12.6 Hz, 1H), 7.64 (d, J = 8.5 Hz, 1H), 4.80-4.58 (m, 1H), 4.42 (s, 1H), 4.15-3.80 (m, 2H), 3.41 (d, J = 25.8 Hz, 2H), 3.00 (d, J = 31.7 Hz, 2H), 2.82 (s, 6H), 2.75 (s, 3H), 1.53 (s, 2H), 1.33 (d, J = 20.5 Hz, 3H), 1.19 (s, 2H). LC-MS: m/z(M + H)⁺ = 533.2. I-22 129

Yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 8.56-8.47 (m, 2H), 8.17 (d, J = 8.5 Hz, 1H), 7.75 (d, J = 11.9 Hz, 1H), 7.47 (d, J = 8.5 Hz, 1H), 3.94 (d, J = 13.4 Hz, 1H), 3.84 (m, 1H), 3.76-3.63 (m, 3H), 3.07-2.85 (m, 10H), 2.65 (m, 3H), 1.95 (s, 9H). LC-MS m/z: (M + H)⁺ = 549.3. I-77 130

Yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 8.56-8.47 (m, 2H), 8.17 (d, J = 8.5 Hz, 1H), 7.75 (d, J = 11.9 Hz, 1H), 7.47 (d, J = 8.5 Hz, 1H), 3.94 (d, J = 13.4 Hz, 1H), 3.84 (m, 1H), 3.76-3.63 (m, 3H), 3.07-2.85 (m, 10H), 2.65 (m, 3H), 1.95 (s, 9H). LC-MS m/z: (M + H)⁺ = 549.3. I-77 131

Yellow solid. 1H NMR (400 MHz, CDCl₃) δ 8.40 (d, J = 3.7 Hz, 1H), 8.26- 8.10 (m, 2H), 7.99 (s, 1H), 7.78 (d, J = 11.9 Hz, 1H), 7.38 (d, J = 8.4 Hz, 1H), 3.69 (q, J = 14.6 Hz, 2H), 3.61-3.52 (m, 1H), 3.30 (t, J = 5.3 Hz, 1H), 3.12 (dd, J = 13.2, 7.4 Hz, 1H), 2.94 (s, 3H), 2.86 (d, J = 5.7 Hz, 1H), 2.74 (d, J = 4.0 Hz, 5H), 2.62 (ddd, J = 33.3, 13.4, 7.3 Hz, 2H), 1.97-1.76 (m, 3H), 1.39-1.29 (m, 3H), 1.25 (s, 2H), 1.13 (t, J = 8.0 Hz, 2H). LC-MS: m/z: (M + H)⁺ = 531.3. I-3 134

Pale yellow solid. 1H NMR (400 MHz, CD₃OD) δ 8.54 (d, J = 3.9 Hz, 1H), 8.49 (s, 1H), 8.33 (s, 1H), 8.19 (d, J = 8.5 Hz, 1H), 7.80 (d, J = 11.8 Hz, 1H), 7.52 (d, J = 8.6 Hz, 1H), 4.94 (s, 1H), 3.76 (s, 2H), 3.38 (d, J = 5.5 Hz, 2H), 3.30- 3.23 (m, 2H), 3.02-2.93 (m, 6H), 2.90 (s, 3H), 2.71 (s, 3H), 1.74 (d, J = 6.9 Hz, 6H), 1.11- 0.99 (m, 2H), 0.11 (s, 9H). LC-MS m/z: (M + H)⁺ = 593. I-3 136

Pale yellow solid. 1H NMR (400 MHz, D₂O) δ 8.34 (s, 1H), 7.81 (d, J = 3.8 Hz, 1H), 7.12 (s, 2H), 6.90 (s, 1H), 6.77 (d, J = 12.1 Hz, 1H), 4.82 (s, 3H), 4.12 (s, 2H), 3.45 (t, J = 6.0 Hz, 2H), 2.85 (s, 2H), 2.20 (s, 3H). LC-MS m/z: (M + H)⁺ = 408.2. I-21 138

Yellow solid. 1H NMR (400 MHz, CD₃OD) δ 8.49 (d, J = 3.8 Hz, 1H), 8.10 (d, J = 7.1 Hz, 1H), 8.00 (s, 1H), 7.75 (d, J = 11.6 Hz, 1H), 7.48 (d, J = 8.4 Hz, 1H), 3.83 (s, 3H), 3.71 (s, 2H), 3.01 (t, J = 6.3 Hz, 2H), 2.94 (s, 4H), 2.85 (t, J = 6.5 Hz, 2H), 2.64 (s, 9H). LC-MS m/z: (M + H)⁺ = 479.3. I-52 139

Yellow solid. 1H NMR (400 MHz, CD₃OD) δ 8.97 (d, J = 3.3 Hz, 1H), 8.56 (s, 1H), 8.23 (dd, J = 16.0, 10.1 Hz, 2H), 7.63 (d, J = 9.0 Hz, 1H), 4.67 (q, J = 7.3 Hz, 2H), 4.52 (s, 2H), 3.75 (t, J = 6.3 Hz, 2H), 3.53 (t, J = 6.1 Hz, 2H), 3.00 (s, 3H), 1.61 (t, J = 7.3 Hz, 3H). LC-MS m/z: (M + H)⁺ = 422.2. I-21 140

Pale yellow solid. 1H NMR (400 MHz, CD₃OD) δ 8.52 (d, J = 3.9 Hz, 1H), 8.13 (d, J = 8.5 Hz, 1H), 8.10 (s, 1H), 7.78 (d, J = 12.0 Hz, 1H), 7.51 (d, J = 8.6 Hz, 1H), 4.37 (q, J = 7.2 Hz, 2H), 3.77 (s, 2H), 3.44-3.39 (m, 2H), 2.97 (m, 12H), 2.69 (s, 3H), 1.48 (t, J = 7.3 Hz, 3H). LC-MS m/z: (M + H)⁺ = 493.3. I-52 141

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 573.2. I-52 142

Pale yellow solid. 1H NMR (400 MHz, DMSO- d₆) δ 9.91 (s, 1H), 8.68 (d, J = 3.9 Hz, 1H), 8.22 (s, 1H), 8.06 (d, J = 8.5 Hz, 1H), 7.75 (d, J = 12.5 Hz, 1H), 7.51 (d, J = 8.4 Hz, 1H), 3.52 (d, J = 40.3 Hz, 4H), 3.33 (s, 17H), 2.75 (dt, J = 70.0, 16.4 Hz, 14H), 2.59- 2.35 (m, 20H), 2.35 (s, 1H), 2.32 (s, 3H), 1.28 (s, 2H), 1.20 (d, J = 53.8 Hz, 3H). LC-MS m/z: (M + H)⁺ = 491.2. I-77 143

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 575.2. I-52 145

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 440.2. Preparation embodi- ment 4 146

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 511.3. I-52 147

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 458.2. Preparation embodi- ment 4 148

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 529.3. I-52 149

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 476.2. Preparation embodi- ment 4 150

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 547.2. I-52 151

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 490.3. Preparation embodi- ment 4 152

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 561.3. I-52 154

Pale yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 8.53 (d, J = 4.0 Hz, 1H), 8.48 (d, J = 1.0 Hz, 1H), 8.43 (s, 1H), 8.20 (d, J = 8.5 Hz, 1H), 7.73 (d, J = 11.6 Hz, 1H), 7.50 (d, J = 8.6 Hz, 1H), 3.93 (s, 2H), 3.15 (dd, J = 17.9, 12.0 Hz, 6H), 3.07-2.97 (m, 5H), 2.94-2.84 (m, 8H), 2.72 (s, 3H), 1.95 (s, 9H). LC-MS m/z: (M + H)⁺ = 576.3. I-52 156

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 490.3. I-52 157

Pale yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 8.51 (d, J = 4.0 Hz, 1H), 8.48 (d, J = 1.2 Hz, 1H), 8.17 (d, J = 8.5 Hz, 1H), 7.73 (d, J = 11.5 Hz, 1H), 7.47 (dd, J = 8.5, 4.5 Hz, 1H), 3.84 (t, J = 5.3 Hz, 2H), 3.79 (s, 1H), 3.71 (dd, J = 10.1, 5.3 Hz, 1H), 3.35 (s, 2H), 3.19-3.14 (m, 2H), 3.04- 2.92 (m, 6H), 2.86 (s, 3H), 2.82 (s, 3H), 1.93 (s, 9H). LC-MS: m/z: (M + H)⁺ = 551.30. I-72 158

Pale yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 8.41 (d, J = 3.8 Hz, 1H), 8.39 (d, J = 1.2 Hz, 1H), 8.20 (d, J = 8.4 Hz, 1H), 8.09 (s, 1H), 7.73 (d, J = 11.4 Hz, 1H), 7.35 (d, J = 8.5 Hz, 1H), 3.66 (s, 2H), 2.95 (t, J = 6.0 Hz, 2H), 2.89 (d, J = 5.6 Hz, 2H), 2.86 (s, 3H), 2.77-2.67 (m, 6H), 2.67- 2.61 (m, 2H), 2.40 (s, 6H), 2.36 (s, 3H), 1.90 (s, 9H). LC-MS m/z: (M + H)⁺ = 578.40. I-153 159

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 611.2. I-3 160

Pale yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 8.51 (d, J = 4.0 Hz, 1H), 8.48 (d, J = 1.0 Hz, 1H), 8.43 (s, 1H), 8.16 (d, J = 8.5 Hz, 1H), 7.73 (d, J = 11.7 Hz, 1H), 7.47 (d, J = 8.6 Hz, 1H), 3.74 (s, 2H), 3.39 (t, J = 5.6 Hz, 2H), 3.31-3.23 (m, 2H), 2.96 (m, 6H), 2.91 (s, 3H), 2.88 (s, 3H), 1.95 (s, 9H), 1.10-1.02 (m, 2H), 0.11 (s, 9H). LC-MS m/z: (M + H)⁺ = 607.3. I-72 161

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 627.2. I-52 162

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 591.3. I-52 163

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 462.3. Preparation embodi- ment4 164

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 533.3. I-52 165

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 464.2. Preparation embodi- ment4 166

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 535.2. I-52 167

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 466.2. Preparation embodi- ment4 168

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 537.2. I-52 169

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 484.2. Preparation embodi- ment4 170

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 555.2. I-52 171

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 450.2. Preparation embodi- ment4 172

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 521.2. I-52 173

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 463.2. Preparation embodi- ment4 174

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 534.2. I-52 175

Yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 8.51 (d, J = 3.9 Hz, 1H), 8.31 (d, J = 1.3 Hz, 1H), 8.16 (d, J = 8.5 Hz, 1H), 7.74 (d, J = 12.0 Hz, 1H), 7.46 (d, J = 8.6 Hz, 1H), 4.94-4.87 (m, 1H), 3.66 (s, 2H), 3.44 (t, J = 6.7 Hz, 2H), 2.93 (d, J = 4.9 Hz, 2H), 2.90 (d, J = 5.0 Hz, 2H), 2.70 (d, J = 6.6 Hz, 2H), 2.68 (s, 3H), 1.95 (s, 3H), 1.72 (s, 3H), 1.70 (s, 3H). LC-MS m/z: (M + H)⁺ = 520.3. I-3 176

Pale yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 8.47 (d, J = 4.2 Hz, 1H), 8.36 (s, 1H), 8.01 (d, J = 8.4 Hz, 1H), 7.52 (d, J = 11.5 Hz, 1H), 7.29-7.21 (m, 1H), 3.52 (s, 2H), 2.80 (d, J = 12.8 Hz, 9H), 2.66 (d, J = 6.6 Hz, 2H), 2.44 (s, 3H), 1.87 (s, 9H). LC-MS m/z: (M + H)⁺ = 507.3. I-77 177

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 590.3. I-95 178

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 547.3. I-72 179

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 545.3. I-72 180

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 579.3. I-52 181

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 611.2. I-187 182

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 609.3. I-187 183

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 610.3. I-187 185

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 598.3. I-187 186

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 612.3. I-187 188

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 545.3. I-52 189

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 549.3. I-52 190

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 535.3. I-52 191

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 535.3. I-52 192

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 549.3. I-52 193

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 563.3. I-52 194

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 563.3. I-52 195

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 643.3. I-3 196

Pale yellow solid. ¹H NMR (400 MHz, CDCl₃)δ 8.46 (br.s, 1H), 8.41 (d, J = 3.9 Hz, 1H), 8.35 (d, J = 1.3 Hz, 1H), 8.17 (d, J = 8.4 Hz, 1H), 7.70 (d, J = 11.4 Hz, 1H), 7.29 (d, J = 8.5 Hz, 1H), 4.19 (pd, J = 7.2, 3.4 Hz, 4H), 3.65 (s, 2H), 3.24 (q, J = 7.6 Hz, 1H), 3.17- 3.11 (m, 1H), 3.07 (q, J = 7.3 Hz, 1H), 3.01 (dd, J = 11.0, 8.3 Hz, 1H), 2.97-2.86 (m, 6H), 2.84 (s, 3H), 2.79 (t, J = 6.6 Hz, 2H), 2.66 (q, J = 8.2 Hz, 1H), 2.51-2.39 (m, 1H), 2.09-1.99. (m, 1H), 1.88 (s, 9H), 1.32 (t, J = 7.1 Hz, 6H). I-153 LC-MS m/z: (M + H)⁺ = 699.35. 198

Pale yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 8.53 (d, J = 4.3 Hz, 1H), 8.40 (s, 1H), 8.05 (d, J = 8.4 Hz, 1H), 7.55 (d, J = 11.7 Hz, 1H), 7.28 (d, J = 8.5 Hz, 1H), 4.21- 4.05 (m, 4H), 3.54 (s, 2H), 2.90- 2.73 (m, 9H), 2.66 (s, 4H), 2.33 (s, 3H), 2.13-2.02 (m, 2H), 1.90 (s, 9H), 1.34 (t, J = 7.1 Hz, 6H). LC-MS m/z: (M + H)⁺ = 671.3. I-72 199

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 522.3. I-137 200

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 550.3. I-137 202

Pale yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 8.50-8.40 (m, 3H), 8.22 (d, J = 8.3 Hz, 1H), 7.75 (d, J = 11.6 Hz, 1H), 7.45 (d, J = 8.7 Hz, 1H), 3.71 (s, 2H), 3.14 (t, J = 5.6 Hz, 2H), 3.01-2.91 (m, 4H), 2.86 (d, J = 6.7 Hz, 5H), 1.94 (s, 9H). LC-MS m/z: (M + H)⁺ = 536. I-71 203

Pale yellow solid. 1H NMR (400 MHz, CD₃OD) δ 8.50 (d, J = 4.2 Hz, 1H), 8.13 (s, 1H), 8.01 (d, J = 8.4 Hz, 1H), 7.51 (d, J = 12.0 Hz, 1H), 7.28 (d, J = 8.4 Hz, 1H), 3.54 (s, 2H), 2.81 (dd, J = 16.5, 5.2 Hz, 6H), 2.72-2.64 (m, 5H), 2.45 (s, 3H), 1.57 (s, 3H), 1.25 (t, J = 6.3 Hz, 2H), 1.20 (t, J = 5.6 Hz, 2H). LC-MS m/z: (M + H)⁺ = 505. I-71 206

Pale yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 8.43 (d, J = 4.5 Hz, 1H), 8.24 (s, 1H), 7.86 (d, J = 8.3 Hz, 1H), 7.31 (d, J = 11.6 Hz, 1H), 7.04 (d, J = 8.3 Hz, 1H), 3.52- 3.34 (m, 3H), 2.79 (s, 3H), 2.74- 2.61 (m, 11H), 2.46-2.39 (m, 1H), 1.81 (s, 9H), 1.21 (d, J = 6.5 Hz, 3H). LC-MS m/z: (M + H)⁺ = 535.40. I-3 207

1H-NMR(400 MHz, CD₃OD) δ = 8.53 (d, J = 4.0 Hz, 1H), 8.50 (d, J = 0.8 Hz, 1H), 8.41 (s, 1H), 8.19 (d, J = 8.4 Hz, 1H), 7.77 (d, J = 12.0 Hz, 1H), 7.50 (d, J = 8.4 Hz, 1H), 6.12 (s, 1H), 3.83 (s, 2H), 3.78 (s, 2H), 3.40-3.32 (m, 4H), 3.02- 3.00 (m, 6H), 2.89 (s, 3H), 2.47- 2.53 (m, 2H), 1.96 (s, 9H). LC-MS: m/z: (M + H)+ = 603.4. I-153 208

Pale yellow solid. 1H NMR (400 MHz, DMSO- d6) δ9.95 (s, 1H), 8.68 (s, 1H), 8.46 (s, 1H), 8.08 (d, J = 8.3 Hz, 1H), 7.69 (d, J = 11.4 Hz, 1H), 7.46 (d, J = 8.0 Hz, 1H), 3.64 (s, 2H), 3.39 (s, 3H), 3.03 (s, 3H), 2.84 (d, J = 6.7 Hz, 8H), 1.88 (s, 9H). LC-MS m/z: (M + H)+ = 556.2. I-3 209

Pale yellow solid. 1H NMR (500 MHz, DMSO- d6) δ10.80 (s, 1H), 9.98 (d, J = 4.9 Hz, 1H), 8.67 (d, J = 3.8 Hz, 1H), 8.45 (s, 1H), 8.13-7.97 (m, 1H), 7.67 (d, J = 11.6 Hz, 1H), 7.52-7.35 (m, 2H), 3.58 (d, J = 15.6 Hz, 2H), 3.41 (d, J = 4.2 Hz, 1H), 3.24 (d, J = 6.1 Hz, 1H), 2.87-2.77 (m, 7H), 1.86 (s, 9H). LC-MS: 507.2(M + H)⁺. I-95 210

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 521.2. I-77 211

Pale yellow solid. LC-MS m/z: (M + H)⁺ = 605.2. I-207

Effect Embodiment 1

Test Method:

CDK kinase inhibitory activity assay used LANCE® Ultra technique, test compounds were screened on CDK4/CycD3, CDK6/CycD3 and CDK2/CycA2 kinases at ATP concentration of Km. During the test, the initial concentration of the test compound was 3333 nM, each test compound set 10 serially diluted concentrations, the dilution fold was 3-fold, each concentration set 2 duplicated wells.

CDK4/CycD3, CDK6/CycD3 and CDK2/CycA2 were purchased from Carna Biosciences, Inc.; dimethyl sulfoxide, ATP, DTT solution were purchased from Sigma-Aldrich; EDTA solution was purchased from GIBCO; LANCE® Detection Buffer, 10× and LANCE® Ultra Europium-anti-phospho-eIF4E-binding protein 1 (Thr37/46) and LANCE® Ultra ULight™-eIF4E-bindingprotein 1 (Thr37/46) Peptide were purchased from Perkinelmer.

Test Procedure:

1. Take 10 mM stock solution of the test compound, in 96-well compound plate, DMSO was used to dilute the compound to an initial concentration of 100×, then this concentration was used as the first concentration, 3-fold diluted to make 10 serial concentrations; 1 μL each serial dilution was then added to 19 μL 1×reaction buffer to prepare 5×compound for use; 2 μL 5×compound was transferred from 96-well plate to 384-well plate; compound-free control well was added with 2 μL the following liquid: 1×reaction buffer with the addition of 1 μL DMSO; 2 μL 250 mM EDTA was added to the Min control well.

2.1× reaction buffer was used to formulate the kinase, substrate and ATP into a 2.5×enzyme/substrate mixture and 2.5×ATP solution respectively. In the experiment, the final concentration of CDK4/CycD3 kinase is 0.76 ng/μL, the final concentration of ATP is 80 μM; the final concentration of CDK6/CycD3 kinase is 0.5 ng/μL, the final concentration of ATP is 50 μM; the final concentration of CDK2/CycA2 kinase is 0.86 ng/μL, the final concentration of ATP is 15 μM; the final concentration of CDK2/CycE1 kinase is 1.016 ng/μL, the final concentration of ATP is 20 μM; 2.5×enzyme/substrate mixture was added to a 384-well plate, incubated at room temperature for 5 minutes; then added with 2.5×ATP solution, reacted at room temperature for 30 minutes.

3. LANCE® Detection Buffer was used, 1× to prepare 2×LANCE® Ultra Europium-anti-phospho-eIF4E-binding protein 1 (Thr37/46) for use. After the enzymatic reaction was continued for 30 minutes, 10 mM EDTA was added to 384-well plate and the mixture was reacted at room temperature for 5 minutes. Then LANCE® Ultra Europium-anti-phospho-eIF4E-binding protein 1 (Thr37/46) was added, reacted at room temperature for 1 hour.

4. The 384-well plate was placed in HERAEUS Multifuge X1R centrifuge, centrifuged at 2000 rpm for 2 minutes; data were measured on EnVision™, 337 nM wavelength laser was used as the excitation light, test at RFU665 nM and RFU615 nM, and RFU665 nM/RFU615 nM×10000 was used as the final data for analysis.

5. Graphpad Prism 5.0 was used to perform Log (inhibitor) vs. response-Variable slope (four parameters) curve fitting on the data and the corresponding IC₅₀ (half maximal inhibitory concentration) was calculated.

The test result was shown in Table 3.

TABLE 2 Structures of comparative examples A-F and LY2835219 comparative example A

comparative example B

comparative example C

comparative example D

comparative example E

comparative example F

LY2835219

TABLE 3 Activity test results on Kinase level Compound CDK4 CDK6 CDK2 number IC₅₀, nM IC₅₀, nM IC₅₀, nM LY2835219 2.51 11.7 52.2 I-3 2.08 10.4 35.0 I-4 1.72 3.49 29.6 I-5 1.64 4.14 33.7 I-6 1.72 7.57 28.7 I-8 2.05 7.07 15.5 I-9 1.74 7.09 13.4 I-10 2.21 7.81 10.4 I-12 3.55 14.2 60.4 I-13 42.3 327 558 I-14 14 140 1090 I-15 20.1 113 306 I-17 2.54 4.88 37.2 I-19 2.34 12.4 38.2 I-20 4.49 22.3 7.34 I-21 4.84 25.0 128 I-22 3.49 10.6 193 I-23 2.88 4.17 14.8 I-24 3.90 6.44 31.1 I-25 3.08 5.78 57.9 I-26 2.23 8.12 37.2 I-27 5.23 14.1 151 I-28 4.11 18.9 143 I-29 1.81 4.98 304 I-30 2.23 3.38 154 I-31 2.35 4.84 20.4 I-32 3.54 13.2 146 I-33 1.65 6.23 65.5 I-34 2.07 2.56 89.7 I-35 1.97 4.53 87.6 I-36 2.75 7.41 281 I-37 4.68 16.1 413 I-38 2.34 4.37 32.3 I-39 2.18 7.43 72.1 I-40 2.53 3.59 40.6 I-41 74.9 272 >3330 I-42 7.05 13.4 84.3 I-43 2.94 3.57 122 I-44 3.21 2.82 54.1 I-45 7.14 22.7 710 I-46 2.52 3.42 346 I-47 2.36 4.16 559 I-48 1.65 4.88 91.0 I-49 3.36 5.17 28.2 I-50 2.71 3.94 334 I-51 1.83 2.88 129 I-52 3.88 3.81 700 I-53 1.70 5.46 314 I-54 4.27 7.50 294 I-55 2.47 3.56 58.9 I-57 3.55 6.14 109 I-58 3.20 6.14 93.7 I-59 3.14 4.78 93.4 I-60 1.47 3.47 79.4 I-61 1.12 2.55 364 I-62 1.44 4.51 28.3 I-63 1.12 2.0 47.7 I-64 3.26 8.47 >1020 I-65 1.35 3.75 >3470 I-66 1.06 6.36 661 I-67 1.53 6.24 212 I-68 1.09 2.97 18.5 I-69 1.13 1.57 54.9 I-70 1.68 5.51 1490 I-71 1.34 3.47 167 I-72 1.12 2.58 110 I-73 3.14 6.91 371 I-74 2.08 3.15 317 I-75 3.37 5.16 74.9 I-76 2.45 4.58 33.9 I-77 2.07 3.15 249 I-78 4.41 5.60 567 I-79 1.92 2.69 35.5 I-80 2.22 3.67 647 I-81 1.68 3.34 207 I-82 0.86 3.16 16.5 I-83 1.08 1.55 17.4 I-84 1.54 3.21 15.0 I-85 0.94 2.54 135 I-86 0.96 1.49 16.3 I-87 0.92 2.21 10.3 I-88 0.91 1.41 16.3 I-89 3.74 17.9 62.4 I-90 1.16 1.56 28.5 I-91 0.99 1.70 16.3 I-92 0.99 1.39 14.3 I-93 0.96 1.33 16.3 I-94 1.16 1.37 10.3 I-95 1.18 2.02 6.02 I-97 1.73 2.11 15.0 I-98 1.04 5.01 670 I-101 1.79 2.90 14.5 I-102 1.02 2.41 19.2 I-103 1.02 0.97 23.8 I-104 4.21 9.25 31.1 I-106 1.33 2.67 83.3 I-107 1.56 4.59 135 I-108 1.61 2.23 116 I-109 1.59 3.86 30.7 I-110 2.00 5.89 14.5 I-111 1.09 2.98 11.2 I-112 1.37 1.70 44.9 I-114 1.74 1.77 23.6 I-126 2.21 6.26 232 I-127 1.65 4.88 444 I-129 0.70 1.13 62.4 I-130 1.02 1.48 14.3 I-131 0.91 1.41 28.5 I-132 0.99 0.77 19.4 I-133 1.52 1.82 78.2 I-134 1.35 1.59 25.1 I-135 17.7 111 147 I-136 14.1 84.2 486 I-137 1.55 4.78 8.43 I-138 4.26 33.0 >3330 I-139 8.12 29.3 126 I-140 2.35 5.67 358 I-142 1.96 3.25 301 I-153 2.26 1.87 42.8 I-154 1.9 1.97 39.6 I-155 1.45 2.08 67.0 I-157 2.06 2.03 48.4 I-158 1.27 1.60 73.1 I-160 3.77 2.46 34.6 I-175 1.86 3.86 21.0 I-176 1.49 1.51 6.68 I-192 1.54 1.53 29.0 I-196 3.06 2.99 60.2 I-197 15.9 12.9 27.6 I-198 1.91 2.03 59.3 I-199 1.53 2.31 27.0 I-200 2.15 5.89 40.2 I-202 1.14 1.11 20.8 I-203 1.51 1.28 20.0 I-204 12.1 25.7 1250 I-205 4.83 14.7 117 I-206 1.56 1.76 50.3 I-207 2.27 1.73 32.0 I-208 1.39 2.58 13.1 I-209 1.48 2.39 14.0 I-210 2.11 1.80 18.0 I-211 1.45 1.36 18.5 comparative 3.80 13.2 17.0 example A comparative 2.93 7.58 13.5 example B comparative 2.37 7.26 21.5 example C comparative 2.06 7.22 31.9 example D comparative 2.06 9.17 19.9 example E comparative 1.72 2.67 22.5 example F

Effect Embodiment 2

MCF-7, T-47D, ZR-75-1, COLO 205 and A549 cell proliferation inhibition assays were performed using the CellTiter-Glo® Luminescent Cell Viability Assay method. Experiments were performed on human breast cancer MCF-7, T-47D and ZR-75-1 cell, human colon cancer COLO 205 cell, and human non-small cell carcinoma A549 cell, used CellTiter method to determine the inhibition effect of the compound on proliferation of five cell lines. During the test, the initial concentration of the test compound was 10M, all test compounds set 10 serially diluted concentrations, the dilution-fold was 3-fold, each concentration set 2 duplicated wells.

Mcf-7 cell, purchased from ATCC, article No. HTB-22;

T-47D, purchased from ATCC, article No. HTB-133;

ZR-75-1, purchased from Shanghai Cell Bank, article No. TCHul26;

COLO 205 human colon cancer cell, purchased from Cell Bank of Type Culture Collection of the Chinese Academy of Sciences/Cell BankShanghai Institutes for Biological Sciences, Chinese Academy of Sciences, catalogue No. TCHul02;

A549 human non-small cell cancer cell, purchased from ATCC, catalogue No. CCL-185™;

CellTiter-Glo®luminescent cell viability assay kit, purchased from Promega, Catalog No. G7573;

RPMI-1640, purchased from Life technologies, article No. A1049101;

F-12K Nutrient Mixture, purchased from Life technologies, article No. 21127-002;

FBS, purchased from Life technologies, article No. 10099-141;

Antibiotic, purchased from Life technologies, article No. 10378-016;

PBS, purchased from Life technologies, article No. 10010-023;

Dimethyl sulfoxide, purchased from Sigma-Aldrich, article No. D8418;

384-well cell culture plate, purchased from Corning, article No. 3570.

Test Procedure:

1. Normally cultured human breast cancer cells, normally cultured 12th generation COLO 205 human colon cancer cells, and normally cultured 7th generation A549 cells were plated in 384-well plate at a certain cell density; MCF-7 and T-47D cells were plated at a density of 500 cells/well, ZR-75-1, COLO 205 cells and A549 cells were plated at a density of 250 cells/well, the cell culture plate was placed at 37° C. in 5% CO₂ condition for 24 hours.

2. One day after human breast cancer cells were plated, one plate was used to measure the background Celltiter value which was recorded as Control D1. Dosing on the remaining plates to set cell control group. The initial concentration of the test compound was 10 μM, followed by 10 serial dilutions, the dilution-fold was 3-fold, each concentration set 2 duplicated well.

2′. After COLO 205 cells and A549 cells were plated for 24 hours, the configured concentration gradient of drug was added to the corresponding cell wells, respectively. The control group Control was a drug-free, medium solution containing 0.2% DMSO. The untreated cell well was used to measure the background value which was recorded as Blank. The cell culture plate was placed at 37° C., COLO 205 cells were cultured in 5% CO₂ condition for 6 days, A549 cells were cultured in 5% CO₂ condition for 4 days.

3. Human breast cancer cells: After 6 days treatment of the test compound, 50 μL CellTiter detection solution was added to each well, the mixture was shaken for 2 min, mixed thoroughly, centrifuged, left to stand for 10 min, the fluorescent signal was recorded. The drug group was recorded as Drug D7. The control group was recorded as Control D7.

3′. COLO 205 cells and A549 cells: the microplate was taken out from incubator and equilibrated at room temperature for 30 minutes. 50 μL CellTiter-Glo® Luminescent Cell Viability Assay which had been equilibrated at room temperature was added to each well, the mixture was shaken at 700 rpm at room temperature for 2 minutes, the microplate was placed in HERAEUS Multifuge X1R centrifuge at 2000 rpm for 1 minute; equilibrated at room temperature for 10 minutes, the fluorescence signal value was measured on EnVision™.

4. Graphpad Prism 5.0 was used to perform Log (inhibitor) vs. response-Variable slope (four parameters) curve fitting on the data and the corresponding IC₅₀ (half maximal inhibitory concentration) was calculated.

The test results were shown in Table 4, wherein the structures of comparative examples A-F refer to those recited in effect embodiment 1.

TABLE 4 Activity results at Cell level COLO MCF-7 T-47D ZR-75-1 205 A549 Compound IC₅₀, IC₅₀, IC₅₀, IC₅₀, IC₅₀, number nM nM nM nM nM LY2835219 116. 38.4 108 411 436 I-3 82.1 79.3 110 213 I-4 185 267 85.2 103 I-5 69.4 9.02 78.4 121 I-6 119 322 253 I-8 154 110 63.2 267 I-9 241 418 176 I-10 895 8680 838 I-12 138 141 1390 I-13 3520 2880 4940 I-14 1590 1370 2230 I-15 >33300 >33300 4760 I-17 176 330 314 I-19 533 198 426 I-20 2090 985 3390 I-21 684 809 335 I-22 245 116 324 I-23 108 20.7 187 120 I-24 93.5 464 235 I-25 96.3 39.4 139 235 I-26 145 637 301 I-27 275 122 1260 I-28 525 212 381 I-29 28.0 16.9 50.0 352 I-30 188 229 1010 I-31 218 660 369 I-32 457 4930 605 I-33 648 1320 269 I-34 116 131 141 I-35 92.3 66.0 73.9 1150 I-36 140 32.9 72.5 542 I-37 666 243 409 1150 I-38 94.1 137 219 161 I-39 2490 137 146 816 I-40 1090 484 369 303 I-41 1320 1430 1500 786 I-42 1050 1200 354 575 I-43 1590 26.1 60.5 598 I-44 771 410 126 367 I-45 2810 283 352 1340 I-46 15.8 47.0 160 522 I-47 35.0 58.8 62.1 637 I-48 27.9 32.5 119 1210 I-49 1590 396 220 195 I-50 88.0 34.6 68.8 614 I-51 314 42.0 83.4 461 I-52 23.3 19.8 25.5 563 49.3 I-53 73.2 23.4 57.0 468 317 I-54 94.1 30.3 56.2 573 I-55 377 16.4 82.5 360 I-57 214 1120 210 478 I-58 431 56.0 88.9 485 I-59 62.3 44.0 35.8 533 I-60 36.2 24.3 61.4 521 I-61 41.5 13.1 68.6 361 I-62 22.6 21.9 49.9 194 I-63 7.03 8.62 21.5 94.5 75.3 I-64 443 72.7 244 3600 I-65 139 33.0 109 72.9 I-66 165 48.1 142 1340 I-67 182 43.8 74.3 607 465 I-68 73.8 131 75.3 96.1 I-69 18.7 5.05 14.1 56.0 58.7 I-70 102 56.7 139 1420 125 I-71 609 2000 692 871 I-72 247 797 130 219 I-73 344 21.3 184 1270 I-74 21.0 8.05 36.1 302 44.1 I-75 308 64.6 272 463 I-76 461 197 232 375 I-77 163 26.9 61.4 541 I-78 136 87.9 164 803 I-79 39.4 14.8 53.6 202 I-80 32.7 42.0 69.6 431 30.8 I-81 145 29.9 60.0 449 I-82 27.4 130 65.7 150 I-83 12.6 8.26 13.6 68.0 179 I-84 22.2 73.9 52.0 93.2 I-85 69.1 16.3 174 250 69.0 I-86 19.9 7.81 29.3 84.0 93.1 I-87 98.4 189 97.0 130 72.6 I-88 21.0 6.29 16.8 70.8 93.3 I-89 522 738 492 770 573 I-90 27.9 8.99 14.5 105 140 I-91 1940 336 1300 3530 3130 I-92 115 18.5 80.0 78.3 69.9 I-93 44.6 14.0 141 189 441 I-94 48.9 25.1 114 335 376 I-95 56.4 179 85.7 114 226 I-97 1380 375 1610 2690 6830 I-98 49.3 47.7 191 578 612 I-101 31.0 61.9 50.7 89.8 54.4 I-102 54.0 62.8 141 1130 1510 I-103 6.99 5.21 16.3 50.4 139 I-104 71.6 237 162 199 148 I-106 23.4 34.7 123 170 337 I-107 551 29.0 110 387 934 I-108 239 118 674 1730 2190 I-109 28.1 94.3 76.7 157 74.7 I-110 152 344 353 350 451 I-111 19.4 77.9 46.4 89.4 70.0 I-112 21.1 4.69 15.0 44.2 195 I-114 7.73 3.24 9.65 40.7 259 I-126 21.2 26.2 61.8 284 68.5 I-127 27.9 32.5 118 1210 I-129 33.1 9.18 133 299 353 I-130 47.14 9.04 45.10 572.50 454 I-131 44.5 30.5 77.2 762 1360 I-132 39.6 405 731 >10000 >10000 I-133 882 810 1360 6210 2690 I-134 86.2 750 479 189 352 I-135 963 1680 1260 1310 1290 I-136 1080 1670 1420 1870 1550 I-137 185.90 140.40 268.40 386.20 482 I-138 224 188 608 2290 3570 I-139 756 808 452 929 1180 I-140 23.2 15.2 80.5 384 2900 I-142 224 15.0 57.9 265 816 I-153 14.8 333.6 41.5 80.6 169 I-154 72.9 9.89 31.3 68.3 I-155 157 106 40.2 425 I-157 296 7.20 19.4 42.2 113 I-158 76.0 45.0 82.5 546 I-160 118 70.5 438 240 424 I-175 612 74.4 261 117 305 I-176 238 5.91 40.4 11.4 37.2 I-187 454 340 574 660 1100 I-192 17.4 10.6 17.7 I-196 51.7 1500 149 425 I-197 957 1240 >10000 7620 >10000 I-198 95.6 26.6 114 96.0 213 I-199 135 169 92.0 I-200 399 76.6 132 125 422 I-202 158 36.0 122 59.4 52.0 I-203 29.8 4.25 14.9 26.1 205 I-204 568 555 1930 5830 >10000 I-205 150 140 452 1060 I-206 20.5 17.8 35.2 I-207 67.2 484 74.8 I-208 66.8 182 56.0 I-209 86.2 127 76.5 I-210 54.6 28.2 72.8 I-211 88.7 229 56.1 comparative 79.1 129 108 114 example A comparative 134 229 309 114 example B comparative 148 119 75.1 371 example C comparative 152 406 367 example D comparative 176 134 59.3 132 example E comparative 291 740 178 example F

Effect Embodiment 3

In vitro proliferation inhibition experiments of U87, MGU251, K562, THP1, HepG2, SK-HEP-1, SNU-5, N87, H1975, H1993, CFPAC1, PANC-1, LNCap, and PC-3 cell lines was used CellTiter-Go® luminescent cell viability assay method. Experiments were performed on U87, MGU251, K562, THP1, HepG2, SK-HEP-1, SNU-5, N87, H1975, H1993, CFPAC1, PANC-1, LNCap and PC-3 cells, CellTiter method was used to determine the inhibition effect of the compound on proliferation. During the test, the initial concentration of the test compound was 10 μM, each test compound set 10 serially diluted concentrations, the dilution-fold was 3-fold, each concentration set 2 duplicated wells.

U87MG human brain astrocytoma cell line, purchased from ATCC, article No. HTB-14

K-562 cell, purchased from Cell Bank of Type Culture Collection of the Chinese Academy of Sciences, article No. TCHu191;

PANC-1 cell, purchased from Cell Bank of Type Culture Collection of the Chinese Academy of Sciences, article No. TCHu98;

THP1 cell, purchased from ATCC, article No. TIB-202;

HepG2 human hepatoma cell line, purchased from ATCC, article No. HB-8065;

SK-HEP-1 cell, purchased from Cell Bank of Type Culture Collection of the Chinese Academy of Sciences, article No. TCHul09;

SNU-5 cell, purchased from ATCC, article No. CRL-5973;

N87 cell, purchased from ATCC, article No. CRL-5822;

H1975 non-small cell lung cancer cell, purchased from ATCC, article No. CRL-5908;

H1993 cell, purchased from ATCC, article No. CRL-5909;

CFPAC1 cell, purchased from ATCC, article No. CRL-1918;

MGU251 cell, purchased from Cell Bank of Type Culture Collection of the Chinese Academy of Sciences, article No. TCHu58;

LNCap cell, purchased from ATCC, article No. CRL-1740;

PC-3 cell, purchased from ATCC, article No. TCHu158;

RPMI-1640 medium, purchased from Life technologies, article No. A1049101;

IMDM medium, purchased from Life technologies, article No. 12440;

DMEM medium, purchased from Life technologies, article No. 11995;

MEM medium, purchased from Life technologies, article No. 51200;

FBS, purchased from Life technologies, article No. 10099;

Antibiotic, purchased from Life technologies, article No. 10378;

PBS, purchased from Life technologies, article No. 10010;

P-mercaptoethanol, purchased from Life technologies, article No. 1150809;

F-12K, purchased from Life technologies, article No. 21127022;

HepG2, purchased from ATCC, article No. HB-8065; U87MG, purchased from ATCC, article No. HTB-14; PC-3, purchased from Cell Bank, Shanghai, article No. TCHul58; CellTiter-Glo©luminescent cell viability assay kit, purchased from Promega, Catalog No. G7573; 384-well cell culture plate, purchased from Corning, article No. 3570; 96-well compound plate, purchased from Nunc, article No. 267245.

Test Procedure:

1. U87MG (DMED medium, 11th generation, 12th generation and 13th generation), MGU251 (4th generation), K562 (10th generation, cell plate density 125 cells/well), THP1 (6th generation, 8th generation and 10th generation), HepG2 (DMED medium, 7th generation, 10th generation and 13th generation), SK-HEP-1 (7th generation), SNU-5 (5th generation), N87 (1640 medium, Assay1: 11th generation, Assay2: 12th generation, Assay3: 13th generation), H1975 (RPMI medium, 6th generation, 9th generation, 11th generation), H1993 (8th generation), CFPAC1 (IMDM medium, Assay1: 9th generation, Assay2: 10th generation, Assay3: 12th generation), PANC-1 (5th generation), LNCap (7th generation, 10th generation and 12th generation, cell plate density 625 cells/well), PC-3 (8th generation), plated in 384-well plate at a cell density of 250 cells/well.

2. One day after plated, one plate was used to measure the background Celltiter value which was recorded as Control D1. Dosing on the remaining plates to set cell control group. The initial concentration of the test compound was 10 μM, followed by 10 serial dilutions, the dilution-fold was 3-fold, each concentration set 2 duplicated well.

3. After certain days (U87MG cell for 4 days, MGU251 cell for 6 days, K562 cell for 4 days, THP1 cell for 4 days, HepG2 cell for 4 days, SK-HEP-1 cell for 4 days, SNU-5 cell for 6 days, N87 cell for 4 days, H1975 cell for 4 days, H1993 cell for 6 days, CFPAC1 cell for 4 days, PANC-1 cell for 4 days, LNCap cell for 6 days, PC-3 cell for 6 days) of treatment of the test compound, 50 μL CellTiter detection solution was added to each well, the mixture was shaken for 2 min, mixed thoroughly, centrifuged, left to stand for 10 min, detected and recorded the fluorescent signal. The drug group was recorded as Drug D7. The control group was recorded as Control D7.

4. Graphpad Prism 5.0 was used to perform Log (inhibitor) vs. response-Variable slope (four parameters) curve fitting on the data and the corresponding IC₅₀ (half maximal inhibitory concentration) was calculated.

The test results were shown in Table 5 and Table 6.

TABLE 5 Activity data of I-52 on different tumor cells Cell LY283521 (IC₅₀, nM) I-52 (IC₅₀, nM) line Assay1 Assay2 Assay3 Assay1 Assay2 Assay3 HepG2 246 204 198 35.5 22.6 21.8 U87MG 711 792 614 37.9 35.5 22.2 LNCap 73.2 81.0 97.8 35.6 67.0 37.5 PC-3 5850 11800 >10000 4190 8760 37500 THP1 251 123 187 36.8 34.1 36.6

TABLE 6 Activity data of I-63 on different tumor cells Cell LY283521 (IC₅₀, nM) I-63 (IC₅₀, nM) line Assay1 Assay2 Assay3 Assay1 Assay2 Assay3 U87 711 792 614 198 154 267 MGU251 232 403 246 111 181 114 K562 974 1110 1190 211 334 352 THP1 251 123 187 61.5 40.9 44.8 HepG2 246 204 198 33.3 24.7 21.8 SK-HEP-1 430 401 402 147 108 153 SNU-5 422 460 685 226 206 340 N87 573 592 391 392 530 346 H1975 603 1040 677 346 367 268 H1993 1120 1240 1100 1000 619 663 CFPAC1 508 378 228 2670 178 272 PANC-1 664 658 782 288 373 297 LNCap 73.2 80.9 97. 8 16.1 31.7 20.1 PC-3 5850 11800 8310 541 535 1480

Effect Embodiment 4 (In Vivo Pharmacodynamics of the Compounds on Human Breast Cancer MCF-7 Cell in BALB/c Nude Mice Subcutaneous Xenograft Model)

Experimental Objective:

Evaluate in vivo pharmacodynamics of the test compounds on human breast cancer MCF-7 cell in BALB/c nude mice subcutaneous xenograft model.

Experimental Design:

64 of BALB/c nude mices, 6-8 weeks old, weight 18-22 g, female, provided by Shanghai Sippr-bk Laboratory Animal Ltd., according to Table 7.

TABLE 7 In vivo pharmacodynamics of experimental animals grouping and dosage regimen Dosing Administration Dosing Group N¹ compound Dose volume route frequency 1 8 menstruum — 10 mL/kg PO QD × 19 days 2 8 Test compound  25 10 mL/kg PO QD × 19 I-52 mg/kg days 3 8 Test compound  50 10 mL/kg PO QD × 19 I-52 mg/kg days 4 8 Test compound 100 10 mL/kg PO QD × 19 I-52 mg/kg days 5 8 Test compound  10 10 mL/kg PO QD × 19 I-63 mg/kg days 6 8 Test compound  25 10 mL/kg PO QD × 19 I-63 mg/kg days 7 8 Test compound  50 10 mL/kg PO QD × 19 I-63 mg/kg days 8 8 Test compound 50 10 mL/kg PO QD × 19 LY2835219 mg/kg days Note: N¹: the number of mice per group Dosing volume: 10 μL/g based on the weight of mice. If the weight loss exceeded 15%, the dosage regimen should be adjusted accordingly.

Experiment Route:

1. Cell culture: human breast cancer cell line MCF-7 was cultured in a monolayer in vitro, the culture condition was in accordance with the supplier's technical specification or the references. Passage cells were treated with trypsin-EDTA for conventional digestion twice a week. When the cell saturation was 80%-90% and the quantity reached the requirements, cells were collected, counted, and inoculated.

2. Estrogen tablet inoculation and urination: 2 days before cell inoculation, the β-estradiol sustained-release tablet was inoculated into the left back of each mice. One week after inoculation, the animal urinated 3 times per week, the animal urinated daily if necessary.

3. Tumor cell inoculation: 0.2 mL 10×10⁶ MCF-7 cells was subcutaneously inoculated into the right back of each nude mice (PBS:Matrigel=1:1). When the average tumor volume reached 195 mm³, the compound was administered according to the dosage regimen (see Table 7).

4. Preparation of the test sample, see Table 8.

TABLE 8 Preparation method of the test compounds Concen- Storage Compound Preparation method tration condition vehicle 1, liquid A: Taking 16.6 m/L 1% 4° C., in phosphoric acid, adding water dark to 1000 mL, shaking; 2, liquid B: Taking 7.163 g disodium phosphate, adding water to 100 mL and making it dissolved; 3, mixing 145 mL the liquid A and 55 mL liquid B, shaking; adding 350 mL pure water, mixing, shaking, determining pH with pH meter to give the phosphate buffer (20 mM, pH 2.0); 4, weighing 5 g hydroxyethyl cellulose, taking 500 mL phosphate buffer (20 mM, pH 2.0), mixing under magnetic stirring, storing at 4° C. in dark ready for use. I-52 Weighing 18 mg compound 2.5 mg/mL 4° C., in I-52, being added to 7.2 mL dark 1% HEC in phosphate buffer, mixing until homogeneous I-52 Weighing 36 mg compound 5 mg/mL 4° C., in I-52, being added into 7.2 mL dark 1% HEC in phosphate buffer, mixing until homogeneous I-52 Weighing 72 mg compound 10 mg/mL 4° C., in I-52, being added into 7.2 mL dark 1% HEC in phosphate buffer, mixing until homogeneous I-63 Weighing 7.2 mg compound 1 mg/mL 4° C., in I-52, being added into 7.2 mL dark 1% HEC in phosphate buffer, mixing until homogeneous I-63 Weighing 18 mg compound 2.5 mg/mL 4° C., in I-52, being added into 7.2 mL dark 1% HEC in phosphate buffer, mixing until homogeneous I-63 Weighing 36 mg compound 5 mg/mL 4° C., in I-52, being added into 7.2 mL dark 1% HEC in phosphate buffer, mixing until homogeneous LY2835219 Weighing 36 mg compound 5 mg/mL 4° C., in LY2835219, being added into dark 7.2 mL 1% HEC in phosphate buffer, mixing until homo- geneous Note: It is necessary to gently homogeneously mix the drug before the drug was administered to the animal.

5, Tumor measurement and experimental indicator: The experimental indicator was to examine whether the tumor growth was inhibit, delayed or cured. Tumor diameter was measured twice a week with a vernier caliper. The formula for calculating the tumor volume was V=0.5a×b², a and b denoted the long diameter and short diameter of the tumor respectively. The antitumor activity of the compounds was evaluated by TGI (%) or relative tumor proliferation rate T/C (%). TGI (%) reflected the tumor growth inhibition rate. Calculation for TGI (%): TGI (%)=[(1−(mean tumor volume at the end of dosing of a treatment group−mean tumor volume at the beginning of dosing of the treatment group))/(mean tumor volume at the end of treatment of the solvent control group-mean tumor volume at the beginning of treatment of the solvent control group)]×100%. Relative tumor proliferation rate T/C (%): the calculation formula was as follows: T/C %=T_(RTV)/C_(RTV)×100% (T_(RTV): RTV of treatment group; C_(RTV): RTV of solvent control group). The relative tumor volume (RTV) was calculated based on the result of tumor measurement, the calculation formula was RTV=V_(t)/V₀, wherein V₀ was the mean tumor volume measured at the beginning (d₀) of dosing, and Vt was the mean tumor volume measured at a certain time, T_(RTV) and C_(RTV) took at the same day. At the end of the experiment, the tumor weight was measured and the tumor weight inhibition rate IR (%) was calculated: the calculation formula was IR (%)=(1-Tw/Vw)×100, Tw: mean tumor weight of the treatment group, Vw: mean tumor weight of the solvent control group.

6. Data summary and statistical analysis: The treatment group showed the best treatment effect on the 19th day after dosing at the end of the experiment. Therefore, based on the data, statistical analysis was performed to evaluate the differences between groups. T-test was used for the comparison between two groups, and one-way ANOVA was used for comparison among three groups or multiple groups. If the variance was not homogeneous, the Games-Howell method was used for test. If variance was homogeneous, the Dunnet (2-sided) method was used for analysis. All data analysis were performed with SPSS 19.0.p<0.05 was considered statistically significant.

Experimental Results

1, Comparison between two groups of the treatment group relative to the control group, see Table 9.

TABLE 9 pharmacodynamics of the compound of the inhibition on MCF-8 xenograft tumorgrowth Day 6 Day 13 Day 19 T/C^(a) TGI^(a) T-test T/C TGI T-test T/C TGI T-test Group (%) (%) p^(b) (%) (%) p (%) (%) p Vehicle — — — — — — — — — LY283 68.96 111.13 0.044 46.02 94.99 0.023 36.58 82.66 0.032 5219  50 mg/ kg I-52 69.27 110.04 0.047 38.21 108.73 0.013 33.66 86.41 0.028  25 mg/ kg I-52 66.78 118.92 0.040 35.49 113.46 0.011 23.09 100.18 0.016  50 mg/ kg I-52 63.65 130.17 0.028 29.9 123.36 0.007 19.44 104.95 0.013 100 mg/ kg I-63 66.45 120.15 0.036 45.53 95.84 0.022 39.49 78.82 0.038  10 mg/ kg I-63 68.73 111.98 0.047 31.43 120.67 0.008 19.49 104.88 0.014  25 mg/ kg I-63 34.11 235.8 0.001 14.48 150.43 0.003 9.82 117.45 0.009  50 mg/ kg Note: ^(a)tumor growth inhibition was calculated by T/C and TGI (TGI(%) = [1-(T_(6/13/19)-T₀)/(V_(6/13/19)-V₀)] × 100); ^(b)p-value comparedthe treatment group with the vehicle group using T-test according to tumor volume.

Each treatment group exhibited inhibitory activity on tumor growth at the 6th day, 13th day and 19th day after dosing compared to the control group. At 19th day after dosing, the mean tumor volume of tumor-bearing mice in the solvent control group reached 840 mm³, compared to that, the effect of the test compounds I-52, I-63 and LY2835219 at different doses was statistically significant. The tumor growth inhibition rate (TGI) of compound I-52 at 25, 50 and 100 mg/kg reached 86.41%, 100.18%, and 104.95%, respectively; the tumor weight inhibition rate (IR) was 72.92%, 81.93%, and 86.35%, respectively; The tumor growth inhibition rate (TGI) of compound I-63 at 10, 25 and 50 and mg/kg reached 78.82%, 104.88%, and 117.45%, respectively; the tumor weight inhibition rate (IR) was 60.42%, 83.38%, and 92.26%, respectively; the tumor growth inhibition rate (TGI) of LY2835219 at 50 mg/kg reached 82.66%, the tumor weight inhibition rate (IR) was 56.18%. The T-test indicated that there was a statistically significant difference with between the treatment group and the vehicle group (p<0.05).

2, Comparison between compound I-52 and compound I-63 at 50 mg/kg, see Table 10.

TABLE 10 Comparison of antitumor activity at 50 mg/kg on MCF-7 xenograft tumor model(based on the tumor volume on the 19th day after dosing) Tumor volumn (mm³)^(a) T/C^(b) TGI^(b) Group (19th day) (%) (%) Pvalue p value LY2835219 306 ± 30 36.58 82.66 0.008^(d) — 50 mg/kg I-52 193 ± 26 23.09 100.18 — 0.008^(e) 50 mg/kg I-63 82 ± 7 9.82 117.45 0.015^(d) 0.000^(e) 50 mg/kg Note: ^(a)mean value ± SEM; ^(b)tumor growth inhibition was calculated by T/C and TGI (TGI(%) = [1 − (T₁₉ − T₀)/V₁₉ − V₀)] × 100); c. p-value was calculated based on tumor volume; ^(d)compared to I-52, 50 mg/kggroup; ^(e)compared to LY2835219, 50 mg/kggroup.

The one-way ANOVA or T-test analysis of the two compounds at 50 mg/kg indicated that there was a significant difference in the inhibitory effect of the three compounds at the same dose on this model (p<0.05) (Table 10)

3, Dose-effect analysis of compound I-52 and I-63, see Table 11

TABLE 11 Comparison of anti-tumor activity of compound I-52 and I-63 at different dose on MCF-7 xenograft tumor model (based on the tumor volume on the 19th day after dosing) Tumor volumn (mm³)^(a) T/C^(b) TGI^(b) Group (19th day) (%) (%) P value P value I-52 283 ± 23 33.66 86.41 — 0.004^(e) 25 mg/kg I-52 193 ± 26 23.09 100.18 0.031^(d) 0.570^(e) 50 mg/kg I-52 163 ± 22 19.44 104.95 0.004^(d) — 100 mg/kg I-63 332 ± 32 39.49 78.82 — 0.000^(g) 10 mg/kg I-63 164 ± 16 19.49 104.88 0.002^(f ) 0.003^(g) 25 mg/kg I-63 82 ± 7 9.82 117.45 0.000^(f ) — 50 mg/kg Note: ^(a)mean value ± SEM; ^(b)tumor growth inhibition was calculated by T/C and TGI (TGI(%) = [1 − (T₁₉ − T₀)/(V₁₉ − V₀)] × 100); c. p-value was calculated based on tumor volume; ^(d)compared to I-52, 25 mg/kg group; ^(e)compared to I-52, 100 mg/kg group; ^(f)compared to I-63, 10 mg/kg group; ^(g)compared to I-63, 50 mg/kg group;

When one-way ANOVA analysis was performed on the three dose groups of compound I-52 respectively, the result indicated that at different doses, there was a significant difference in the anti-tumor activity between the low dose group of compound I-52 and the middle, high dose group of compound I-52 (p<0.05), but there was no significant difference between the medium dose group and the high dose group (Table 10).

When one-way ANOVA analysis was performed on the three dose groups of compound I-63 respectively, the result indicated that at different doses, there was a significant difference in the anti-tumor activity of compound I-63 (Table 11).

Effect Embodiment 5

Test Method:

CDK kinase inhibitory activity assay used LANCE® Ultra technique, test compounds were screened on CDK1/CycB, CDK5/p25, CDK7/CycH/MAT1 and CDK9/CycT1 kinases at ATP concentration of Km. During the test, the initial concentration of the test compound was 3333 nM, each test compound set 10 serially diluted concentrations, the dilution-fold was 3-fold, each concentration set 2 duplicated wells.

CDK1/CycB, CDK5/p25 and CDK7/CycH/MAT1 were purchased from Carna Biosciences, Inc.; CDK9/CycT1 was purchased from Lifetech; dimethyl sulfoxide, ATP, DTT solution were purchased from Sigma-Aldrich; EDTA solution was purchased from GIBCO; LANCE® Detection Buffer, 10× and LANCE® Ultra Europium-anti-phospho-eIF4E-binding protein 1 (Thr37/46) and LANCE® Ultra ULight™-eIF4E-bindingprotein 1 (Thr37/46) Peptide were purchased from Perkinelmer.

Test Procedure:

1, Take 10 mM test compound stock solution, in 96-well compound plate, DMSO was used to make compound with an initial concentration of 100×, then this concentration was used as the first concentration, 3-fold diluted to make 10 serial concentrations; 1 μL each serial dilution was then added to 19 μL 1× reaction buffer to prepare 5×compound for use; 2 μL 5×compound was transferred from 96-well plate into 384-well plate; compound-free control well was added 2 μL the following liquid: 19 μL 1× reaction buffer with the addition of 1 μL DMSO; 2 μL 250 mM EDTA was added to the Min control well.

2,1× reaction buffer was used to formulate the kinase, substrate, and ATP into a 2.5×enzyme/substrate mixture and 2.5×ATP solution respectively. In the experiment, the final concentration of CDK1/CycB kinase was 3.20 ng/μL, the final concentration of ATP was 12 μM; the final concentration of CDK5/p25 kinase was 0.0334 ng/μL, the final concentration of ATP was 4 μM; the final concentration of CDK7/CycH/MAT1 kinase was 1.93 ng/μL, the final concentration of ATP was 20 μM; the final concentration of CDK9/CycT1 kinase was 0.60 ng/μL, the final concentration of ATP was 12 μM; added 2.5×enzyme/substrate mixture to a 384-well plate, incubated at room temperature for 5 minutes; then added 2.5×ATP solution, reacted at room temperature for 30 minutes.

3, LANCE® Detection Buffer was used, 1× to prepare 2× LANCE® Ultra Europium-anti-phospho-eIF4E-binding protein 1 (Thr37/46) for use. After the enzymatic reaction was continued for 30 minutes, 10 mM EDTA was added to 384-well plate and reacted at room temperature for 5 minutes. Then LANCE® Ultra Europium-anti-phospho-eIF4E-binding protein 1 (Thr37/46) was added, reacted at room temperature for 1 hour.

4,384-well plate was placed in HERAEUS Multifuge X1R centrifuge, centrifuged at 2000 rpm for 2 minutes; data was measured on EnVision™, 337 nM wavelength laser was selected as the excitation light, measured at RFU665 nM and RFU615 nM, and RFU665 nM/RFU615 nM×10000 was used as the final data for analysis.

5, Graphpad Prism 5.0 was used to perform Log (inhibitor) vs. response-Variable slope (four parameters) curve fitting on the data and the corresponding IC₅₀ (half maximal inhibitory concentration) was calculated.

The test results were shown in Table 12.

TABLE 12 Activity test result at kinase level Compound CDK1 CDK5 CDK7 CDK9 number IC₅₀, nM IC₅₀, nM IC₅₀, nM IC₅₀, nM LY2835219 263.2/557.90 81.80/51.80 667.10/225.90 34.55/49.28 I-5 97.25 913.41 13.04 I-8 73.16 393.28 11.43 I-9 62.25 13156.54 7.52 I-22 183.44 1044.56 36.09 I-23 44.80 801.32 8.21 I-24 61.16 2892.62 8.51 I-25 86.66 978.09 10.54 I-29 222.26 573.79 48.04 I-38 26.21 >30000 0.92 I-46 177.50 1280.02 82.04 I-47 491.11 563.30 104.66 I-50 726.02 718.19 89.84 I-51 119.19 1153.10 65.83 I-52 368.21 91.52 223.30 332.39 I-54 357.21 1817.51 80.43 I-59 583.83 650.34 152.40 I-60 306.33 1194.59 82.12 I-61 347.63 710.25 61.45 I-62 69.04 2818.63 2.44 I-63 143.40 33.57 261.10 30.28 I-65 360.01 1076.14 88.95 I-67 232.37 555.53 26.64 I-68 87.81 41.00 20.47 6.94 I-69 138.80 11.44 129.20 16.22 I-70 3614.00 620.20 196.40 316.20 I-74 911.90 36.99 104.00 211.50 I-79 209.00 82.39 279.80 47.45 I-80 1096.00 91.22 146.60 223.30 I-82 125.5 22.56 11.40 4.97 I-83 171.9 28.99 38.13 12.51 I-84 107.8 42.32 16.70 6.58 I-86 101.9 15.35 60.97 10.68 I-88 283.5 33.29 95.93 27.99 I-90 218.2 40.30 36.48 22.75 I-92 87.39 27.99 50.13 7.47 I-93 199.4 139.60 147.00 13.98 I-94 264.1 67.33 66.25 13.53 I-98 1702.0 288.30 669.70 126.9 I-101 78.94 3.90 56.98 6.74 I-109 164.7 47.24 99.42 7.03 I-111 55.07 15.92 104.8 5.44 I-114 161.2 14.61 195.3 15.32 I-126 291.56 1181.63 47.08 I-127 997.52 779.46 145.43 I-129 132.9 55.27 53.40 10.99 I-130 207.9 85.75 109.10 20.79 I-131 1054 238.60 124.10 163.70 I-142 1233 29.33 314.7 133.1 I-154 442.6 15.57 180.8 20.72 I-157 273.1 21.36 214.6 23.56 I-176 134.6 8.37 148.7 10.54 I-202 141.3 7.17 159.5 8.42 I-203 546.6 28.46 250.1 42.29 comparative 50.78 325.62 9.21 example A comparative 41.12 5366.87 9.18 example B comparative 131.93 1094.50 14.77 example C comparative 83.36 799.36 10.80 example E

It is to be understood that the foregoing description of the preferred embodiments is intended to be purely illustrative of the principles of the invention, rather than exhaustive thereof, and that changes and variations will be apparent to those skilled in the art, and that the present invention is not intended to be limited other than expressly set forth in the following claims. 

What is claimed is:
 1. A compound represented by formula I:

or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof, wherein: m is 1; n is 0; p is 1; q is 1; R¹ is F; R² is CH₃; R³ is CH(CH₃)₂; R⁴ is F; R⁵ is (CH₂)₂N(R^(11a))(R^(11b)); R^(6a) is H; R^(6b) is H; R^(6c) is H; R^(6d) is H; R^(11a) is C₁-C₃ alkyl; R^(11b) is C₁-C₃ alkyl; X is N; and Y is CH.
 2. The compound according to claim 1, or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof, wherein R^(11a) is C₂-C₃ alkyl.
 3. The compound according to claim 1, or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof, wherein R^(11a) is CH₃ or CH₂CH₃.
 4. The compound according to claim 1, or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof, wherein R^(11b) is C₂-C₃ alkyl.
 5. The compound according to claim 1, or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof, wherein R^(11b) is CH₃ or CH₂CH₃.
 6. The compound according to claim 1, or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof, wherein: R^(11a) is C₂-C₃ alkyl; and R^(11b) is C₂-C₃ alkyl.
 7. The compound according to claim 1, or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof, wherein: R^(11a) is CH₃ or CH₂CH₃; and R^(11b) is CH₃ or CH₂CH₃.
 8. The compound according to claim 1, wherein the compound is:

or a pharmaceutically acceptable salt or tautomer thereof.
 9. A pharmaceutical composition comprising at least one pharmaceutically acceptable excipient and a compound according to claim 1, or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof.
 10. A method for inhibiting cyclin-dependent kinase activity in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the compound according to claim 1, or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof.
 11. A method for inhibiting tumor cells in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the compound according to claim 1, or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof.
 12. The method according to claim 11, wherein the tumor is selected from the group consisting of acute monocytic leukemia, brain astrocytoma, breast cancer, chronic myelogenous leukemia, colon cancer, gastric carcinoma, hepatocellular carcinoma, liver adenocarcinoma, malignant glioblastoma, non-small cell carcinoma, non-small cell lung cancer, pancreatic cancer, and prostate adenocarcinoma.
 13. A method for treating a tumor in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the compound according to claim 1, or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof.
 14. The method according to claim 13, wherein the tumor is selected from the group consisting of acute monocytic leukemia, brain astrocytoma, breast cancer, chronic myelogenous leukemia, colon cancer, gastric carcinoma, hepatocellular carcinoma, liver adenocarcinoma, malignant glioblastoma, non-small cell carcinoma, non-small cell lung cancer, pancreatic cancer, and prostate adenocarcinoma.
 15. A compound selected from the group consisting of:

or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof.
 16. A compound selected from the group consisting of:

or a pharmaceutically acceptable salt or tautomer thereof.
 17. A compound selected from the group consisting of:

or a stereoisomer or tautomer thereof.
 18. A compound selected from the group consisting of:

or a tautomer thereof. 